5,6-dihydro-1H-pyridin-2-one compounds

ABSTRACT

The invention is directed to 5,6-dihydro-1H-pyridin-2-one compounds and pharmaceutical compositions containing such compounds that are useful in treating infections by hepatitis C virus.

This application is a continuation of U.S. application Ser. No.14/041,376, filed Sep. 30, 2013, which is a divisional of U.S.application Ser. No. 13/567,667, filed Aug. 6, 2012, issuing as U.S.Pat. No. 8,546,602 on Oct. 1, 2013, which is a divisional of U.S.application Ser. No. 13/356,063, filed Jan. 23, 2012, issued as U.S.Pat. No. 8,236,948 on Aug. 7, 2012, which is a divisional of U.S.application Ser. No. 13/043,167, filed Mar. 8, 2011, issued as U.S. Pat.No. 8,101,800 on Jan. 24, 2012, which is a divisional of U.S.application Ser. No. 12/061,499, filed Apr. 2, 2008, issued as U.S. Pat.No. 7,939,524 on May 10, 2011, which claims the benefit of U.S.Provisional Application No. 60/907,478, filed Apr. 3, 2007, the entirecontents of which are incorporated herein.

FIELD OF THE INVENTION

The invention is directed to 5,6-dihydro-1H-pyridin-2-one compounds andpharmaceutical compositions containing such compounds that are useful intreating infections by hepatitis C virus.

BACKGROUND OF THE INVENTION

Hepatitis C is a major health problem world-wide. The World HealthOrganization estimates that 170 million people are chronic carriers ofthe hepatitis C virus (HCV), with 4 million carriers in the UnitedStates alone. In the United States, HCV infection accounts for 40% ofchronic liver disease and HCV disease is the most common cause for livertransplantation. HCV infection leads to a chronic infection and about70% of persons infected will develop chronic histological changes in theliver (chronic hepatitis) with a 10-40% risk of cirrhosis and anestimated 4% lifetime risk of hepatocellular carcinoma. The CDCestimates that each year in the United States there are 35,000 new casesof HCV infection and approximately ten thousand deaths attributed to HCVdisease.

The current standard of care is a pegylated interferon/ribavirincombination at a cost of approximately $31,000/year. These drugs havedifficult dosing problems and side-effects that preclude their use inalmost half of diagnosed patients. Pegylated interferon treatment isassociated with menacing flu-like symptoms, irritability, inability toconcentrate, suicidal ideation, and leukocytopenia. Ribavirin isassociated with hemolytic anemia and birth defects.

The overall response to this standard therapy is low; approximately onethird of patients do not respond. Of those who do respond, a largefraction relapses within six months of completing 6-12 months oftherapy. As a consequence, the long-term response rate for all patientsentering treatment is only about 50%. The relatively low response rateand the significant side-effects of current therapy anti-HCV drugtreatments, coupled with the negative long term effects of chronic HCVinfection, result in a continuing medical need for improved therapy.Antiviral pharmaceuticals to treat RNA virus diseases like HCV are few,and as described above are often associated with multiple adverseeffects.

A number of publications have described NS5B inhibitors useful in thetreatment of hepatitis C infection. See, e.g., U.S. Patent ApplicationPublication No. US 2008/0031852 (describing [1,2-b]pyridazinonecompounds); U.S. Patent Application Publication No. US 2006/0189602(disclosing certain pyridazinones); U.S. Patent Application PublicationNo. US 2006/0252785 (disclosing selected heterocyclics); andInternational Publication Nos. WO 03/059356, WO 2002/098424, and WO01/85172 (each describing a particular class of substitutedthiadiazines).

While there are, in some cases, medicines available to reduce diseasesymptoms, there are few drugs to effectively inhibit replication of theunderlying virus. The significance and prevalence of RNA virus diseases,including but not limited to chronic infection by the hepatitis C virus,and coupled with the limited availability and effectiveness of currentantiviral pharmaceuticals, have created a compelling and continuing needfor new pharmaceuticals to treat these diseases.

SUMMARY OF THE INVENTION

The present invention describes novel 5,6-dihydro-1H-pyridin-2-onecompounds and pharmaceutically acceptable salts thereof, which areuseful in treating or preventing a hepatitis C virus infection in apatient in need thereof comprising administering to the patient atherapeutically or prophylactically effective amount of a5,6-dihydro-1H-pyridin-2-one compound.

In a general aspect, the invention relates to compounds of Formula I

wherein

-   X is N or CR³,-   A is

Ring B is 6-membered aryl or heterocyclyl, optionally substituted by 1-3R¹ moieties, wherein is R¹ is H, halo, nitro, —CHR⁴—S(O)₂R⁵,—C(S(O)₂R⁵)═CHR⁴—, —NR⁵R⁶, —NR⁴S(O)₂R⁵, or —NR⁴S(O)₂NR⁵R⁶, wherein R⁴,R⁵, and R⁶ are independently H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,C(O)O—(C₁-C₆ alkyl), aryl, or heterocyclyl, or R⁴ and R⁵ or R⁵ and R⁶combine with the atom(s) to which they are attached to form a 5- or6-membered heterocyclyl ring,

-   R² is H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, —C₁-C₆ alkylene(C₃-C₈    cycloalkyl), —C₁-C₆ alkylene(aryl), —C₁-C₆ alkylene(heterocyclyl),    aryl, or heterocyclyl,-   R³ is H, halo, or C₁-C₆ alkyl,-   Z is —(CR¹³R¹⁴)_(n)—, or O,-   n is 1 or 2,-   R⁷ and R⁸ are independently H or C₁-C₆ alkyl,-   R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are independently H, C₁-C₆    alkyl, hydroxy, or halo,

wherein the above alkyl, alkylene, aryl, cycloalkyl, or heterocyclylmoieties provided in R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ are each optionally and independently substitutedby 1-3 substituents selected from

-   -   alkylamine,    -   amino,    -   aryl, cycloalkyl, heterocyclyl,    -   C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,        C₁-C₆    -   alkylamine, C₁-C₆ dialkylamine, C₂-C₆ alkenyl, or C₂-C₆ alkynyl,        wherein each    -   of which may be interrupted by one or more hetero atoms,    -   carboxyl,    -   cyano,    -   halo,    -   hydroxy,    -   keto,    -   nitro,    -   —C(O)OH, —C(O)NH₂, —C(O)(C₁-C₆ alkylamine), —C(O)(C₁-C₆        dialkylamine), —C(O)₂—(C₁-C₆ alkyl), —C(O)₂—(C₃-C₈ cycloalkyl),        —C(O)₂-(aryl), —C(O)₂-(heterocyclyl), —C(O)₂—(C₁-C₆        alkylene)aryl, —C(O)₂—(C₁-C₆ alkylene)heterocyclyl,        —C(O)₂—(C₁-C₆ alkylene)cycloalkyl, —C(O)(C₁-C₆ alkyl),        —C(O)(C₃-C₈ cycloalkyl), —C(O)(aryl), —C(O)(heterocyclyl),        —C(O)(C₁-C₆ alkylene)aryl, —C(O)(C₁-C₆ alkylene)heterocyclyl,        and —C(O)(C₁-C₆ alkylene)cycloalkyl,

wherein each of the above optional substituents can be furtheroptionally substituted by 1-5 substituents selected from amino, cyano,halo, hydroxy, nitro, C₁-C₆ alkylamine, C₁-C₆ dialkylamine, C₁-C₆ alkyl,C₁-C₆ alkoxy, C₁-C₆ alkenyl, and C₁-C₆ hydroxyalkyl, wherein each alkylis optionally substituted by one or more halo substituents,

or a pharmaceutically acceptable salt, hydrate, solvate, tautomer orstereoisomer thereof.

In one embodiment, the invention relates to compounds of Formula Iwherein Ring B is selected from

In another embodiment, Ring B is

wherein R¹ is H, halo, nitro, —CHR⁴—S(O)₂R⁵, —C(S(O)₂R⁵)═CHR⁴—, —NR⁵R⁶,—NR⁴S(O)₂R⁵, or —NR⁴S(O)₂NR⁵R⁶.

In one embodiment, the invention relates to compounds of Formula Iwherein R¹ is —NR⁴S(O)₂R⁵, wherein R⁴ and R⁵ are independently H, C₁-C₆alkyl or C₃-C₈ cycloalkyl.

In another embodiment, the invention relates to compounds of Formula Iwherein R¹ is selected from

In one embodiment, the invention relates to compounds of Formula Iwherein R² is selected from

In another embodiment, the invention relates to compounds of Formula Iwherein R² is selected from

In yet another embodiment, the invention relates to compounds of FormulaI wherein R² is selected from

In one embodiment, the invention relates to compounds of Formula Iwherein R³ is selected from hydrogen or C₁-C₆ alkyl.

In another embodiment, the invention relates to compounds of Formula Iwherein R³ is selected from

In one embodiment, the invention relates to compounds of Formula Iwherein R⁷ and R⁸ are H.

In one embodiment, the invention relates to compounds of Formula Iwherein R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are independentlyselected from

In a further embodiment, the invention relates to compounds of Formula Iwherein R⁹, R¹⁰, R¹¹, R¹², R³, R¹⁴, R¹⁵, and R¹⁶ are independentlyselected from

In another embodiment, the invention relates to compounds of Formula Iwherein R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are H or hydroxy.

In yet another embodiment, the invention relates to compounds of FormulaI wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are H.

In one embodiment n is 1.

In another embodiment, the invention relates to compounds selected from

-   (rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-endo)-N-{3-[3-(5-Fluoro-pyridin-2-ylmethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1S,2S,7R,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2R,7S,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methane    sulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methane    sulfonamide,-   (1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)-5-[7-(1,1-Dioxo-4,5-dihydro-1H-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)-5-[7-(1,1-Dioxo-tetrahydro-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide,-   (rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)-5-[7-(1,1-Dioxo-1λ⁶-isothiazolidin-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)—N-[3-(6-Hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,-   (1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)-5-(7-Amino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   Cyclopropanesulfonic acid    {3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   (rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide,-   Cyclopropanesulfonic acid    {3-(1R,2S,7R,8S)-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   (1R,2S,7R,8S)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-benzenesulfonamide,-   (rac-di-exo)-Cyclopropanesulfonic acid    {3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   (rac-di-exo)-Cyclopropanesulfonic acid    {3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   N-[3-(1R,2S,7R,8S)-3-Cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,-   (rac-di-exo)-Cyclopropanesulfonic acid    {3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   (rac-di-exo)-Cyclopropanesulfonic acid    {3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   N-[3-(1R,2S,7R,8S)-(3-Cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide-N-isopropyl    carbamate,-   (rac-di-exo)-N-[3-(3-Cyclopentyl-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,-   N-{3-[(2S,7R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   cis-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (2R,7S)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tetracyclo[6.3.2.0^(2,7).0^(9,11)]tridec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (2S,7R)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tetracyclo[6.3.2.0^(2,7).0^(9,11)]tridec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)-5-(1,1-Dioxo-7-pyrrolidin-1-yl-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-6-hydroxy-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   Pyridine-3-sulfonic acid    {3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide,-   1R,2S,7R,8S)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamide,-   (1R,2S,7R,8S)—N-[3-(3-Benzyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,-   (1R,2S,7R,8S)—N-[3-(6-Hydroxy-3-isobutyl-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,-   (1R,2S,7R,8S)—N-{3-[3-(3-Chloro-4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methane    sulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(3-Fluoro-4-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)-5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   (1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   (1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[3,2-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,-   N-{3-[(1R,2S,7R,8S)-3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6,9-dihydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,    L-arginine salt,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,    L-lysine salt,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,    hemi magnesium salt,-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,    sodium salt, and-   N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,    potassium salt.

The invention is also directed to pharmaceutically acceptable salts andpharmaceutically acceptable solvates of the compounds of Formula I.Advantageous methods of making the compounds of Formula I are alsodescribed.

In one aspect, the invention encompasses a method for treating orpreventing hepatitis C virus infection in a mammal in need thereof,preferably in a human in need thereof, comprising administering to thepatient a therapeutically or prophylactically effective amount of aFormula I compound. In one embodiment, the invention encompasses amethod for treating or preventing hepatitis C virus infection byadministering to a patient in need thereof a therapeutically orprophylactically effective amount of a Formula I compound that is aninhibitor of HCV NS5B polymerase.

In another aspect, the invention encompasses a method for treating orpreventing hepatitis C virus infection in a patient in need thereof,comprising administering to the patient a therapeutically orprophylactically effective amount of a compound of Formula I and apharmaceutically acceptable excipient, carrier, or vehicle.

In another aspect, the invention encompasses a method for treating orpreventing hepatitis C virus infection in a patient in need thereof,comprising administering to the patient a therapeutically orprophylactically effective amount of a compound of Formula I and anadditional therapeutic agent, preferably an additional antiviral agentor an immunomodulatory agent.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a x-ray diffraction spectrum ofN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(as prepared in Example 6 on a kg scale).

FIG. 2 shows a FT-Raman spectrum ofN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(as prepared in Example 6 on a kg scale).

DETAILED DESCRIPTION OF THE INVENTION

Where the following terms are used in this specification, they are usedas defined below:

The terms “comprising,” “having” and “including” are used herein intheir open, non-limiting sense.

The term “alkyl”, as used herein, unless otherwise indicated, includesC₁-C₁₂ saturated monovalent hydrocarbon radicals having straight,branched, or cyclic moieties (including fused and bridged bicyclic andspirocyclic moieties), or a combination of the foregoing moieties. Foran alkyl group to have cyclic moieties, the group must have at leastthree carbon atoms.

The term “alkylene”, as used herein, unless otherwise indicated,includes a C₁-C₁₂ divalent radical derived from alkyl, as exemplified by—CH₂CH₂CH₂CH₂—.

The term “alkenyl”, as used herein, unless otherwise indicated, includesC₁-C₁₂ alkyl moieties having at least one carbon-carbon double bondwherein alkyl is as defined above and including E and Z isomers of saidalkenyl moiety.

The term “alkynyl”, as used herein, unless otherwise indicated, includesC₁-C₁₂ alkyl moieties having at least one carbon-carbon triple bondwherein alkyl is as defined above.

The term “alkoxy”, as used herein, unless otherwise indicated, includesO-alkyl groups wherein alkyl is as defined above.

The term “Me” means methyl, “Et” means ethyl, and “Ac” means acetyl.

The term “cycloalkyl”, as used herein, unless otherwise indicated refersto a non-aromatic, saturated or partially saturated, monocyclic orfused, spiro or unfused bicyclic or tricyclic hydrocarbon referred toherein containing a total of from 3 to 10 carbon atoms, preferably 5-8ring carbon atoms. Exemplary cycloalkyls include monocyclic rings havingfrom 3-7, preferably 3-6, carbon atoms, such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl and the like. Illustrative examplesof cycloalkyl are derived from, but not limited to, the following:

The term “aryl”, as used herein, unless otherwise indicated, includes anorganic radical derived from an aromatic hydrocarbon by removal of onehydrogen, such as phenyl or naphthyl, containing a total of from 6 to 10carbon atoms.

The term “heterocyclic” or “heterocyclyl”, as used herein, unlessotherwise indicated, includes aromatic (e.g., heteroaryls) andnon-aromatic heterocyclic groups containing one to four heteroatoms eachselected from O, S and N, wherein each heterocyclic group has from 4-10atoms in its ring system, and with the proviso that the ring of saidgroup does not contain two adjacent O atoms. Non-aromatic heterocyclicgroups include groups having only 3 atoms in their ring system, butaromatic heterocyclic groups must have at least 5 atoms in their ringsystem. The heterocyclic groups include benzo-fused ring systems. Anexample of a 4 membered heterocyclic group is azetidinyl (derived fromazetidine). An example of a 5 membered heterocyclic group is thiazolyland an example of a 10 membered heterocyclic group is quinolinyl.Examples of non-aromatic heterocyclic groups are pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl,homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl,indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl,pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl andquinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl,imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl,furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl,furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl,benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, andfuropyridinyl. The foregoing groups, as derived from the groups listedabove, may be C-attached or N-attached where such is possible. Forinstance, a group derived from pyrrole may be pyrrol-1-yl (N-attached)or pyrrol-2-yl (C-attached) or pyrrol-3-yl (C-attached). Further, agroup derived from imidazole may be imidazol-1-yl (N-attached) orimidazol-2-yl (C-attached). The 4-10 membered heterocyclic may beoptionally substituted on any ring carbon, sulfur, or nitrogen atom(s)by one to two oxo, per ring. An example of a heterocyclic group wherein2 ring carbon atoms are substituted with oxo moieties is1,1-dioxo-thiomorpholinyl. Other illustrative examples of 4-10 memberedheterocyclic are derived from, but not limited to, the following:

Unless defined otherwise, “alkyl,” “alkylene,” “alkenyl,” “alkynyl,”“aryl,” “cycloalkyl,” or “heterocyclyl” are each optionally andindependently substituted by 1-3 substituents selected from alkylamine,amino, aryl, cycloalkyl, heterocyclyl, C₁-C₆ alkyl, C₁-C₆ haloalkyl,C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ alkylamine, C₁-C₆ dialkylamine,C₂-C₆ alkenyl, or C₂-C₆ alkynyl, wherein each of which may beinterrupted by one or more hetero atoms, carboxyl, cyano, halo, hydroxy,nitro, —C(O)OH, —C(O)₂—(C₁-C₆ alkyl), —C(O)₂—(C₃-C₈ cycloalkyl),—C(O)₂-(aryl), —C(O)₂-(heterocyclyl), —C(O)₂—(C₁-C₆ alkylene)aryl,—C(O)₂—(C₁-C₆ alkylene)heterocyclyl, —C(O)₂—(C₁-C₆ alkylene)cycloalkyl,—C(O)(C₁-C₆ alkyl), —C(O)(C₃-C₈ cycloalkyl), —C(O)(aryl),—C(O)(heterocyclyl), —C(O)(C₁-C₆ alkylene)aryl, —C(O)(C₁-C₆alkylene)heterocyclyl, and —C(O)(C₁-C₆ alkylene)cycloalkyl, wherein eachof these optional substituents can be further optionally substituted by1-5 substituents selected from amino, cyano, halo, hydroxy, nitro, C₁-C₆alkylamine, C₁-C₆ dialkylamine, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆alkenyl, and C₁-C₆ hydroxyalkyl, wherein each alkyl is optionallysubstituted by one or more halo substituents, e.g., CF₃.

The term “immunomodulator” refers to natural or synthetic productscapable of modifying the normal or aberrant immune system throughstimulation or suppression.

The term “preventing” refers to the ability of a compound or compositionof the invention to prevent a disease identified herein in patientsdiagnosed as having the disease or who are at risk of developing suchdisease. The term also encompasses preventing further progression of thedisease in patients who are already suffering from or have symptoms ofsuch disease.

The term “patient” or “subject” means an animal (e.g., cow, horse,sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit, guineapig, etc.) or a mammal, including chimeric and transgenic animals andmammals. In the treatment or prevention of HCV infection, the term“patient” or “subject” preferably means a monkey or a human, mostpreferably a human. In a specific embodiment the patient or subject isinfected by or exposed to the hepatitis C virus. In certain embodiments,the patient is a human infant (age 0-2), child (age 2-17), adolescent(age 12-17), adult (age 18 and up) or geriatric (age 70 and up) patient.In addition, the patient includes immunocompromised patients such as HIVpositive patients, cancer patients, patients undergoing immunotherapy orchemotherapy. In a particular embodiment, the patient is a healthyindividual, i.e., not displaying symptoms of other viral infections.

The term a “therapeutically effective amount” refers to an amount of thecompound of the invention sufficient to provide a benefit in thetreatment or prevention of viral disease, to delay or minimize symptomsassociated with viral infection or viral-induced disease, or to cure orameliorate the disease or infection or cause thereof. In particular, atherapeutically effective amount means an amount sufficient to provide atherapeutic benefit in vivo. Used in connection with an amount of acompound of the invention, the term preferably encompasses a non-toxicamount that improves overall therapy, reduces or avoids symptoms orcauses of disease, or enhances the therapeutic efficacy of or synergieswith another therapeutic agent.

The term a “prophylactically effective amount” refers to an amount of acompound of the invention or other active ingredient sufficient toresult in the prevention of infection, recurrence or spread of viralinfection. A prophylactically effective amount may refer to an amountsufficient to prevent initial infection or the recurrence or spread ofthe infection or a disease associated with the infection. Used inconnection with an amount of a compound of the invention, the termpreferably encompasses a non-toxic amount that improves overallprophylaxis or enhances the prophylactic efficacy of or synergies withanother prophylactic or therapeutic agent.

The term “in combination” refers to the use of more than oneprophylactic and/or therapeutic agents simultaneously or sequentiallyand in a manner that their respective effects are additive orsynergistic.

The term “treating” refers to:

(i) preventing a disease, disorder, or condition from occurring in ananimal that may be predisposed to the disease, disorder and/orcondition, but has not yet been diagnosed as having it;

(ii) inhibiting the disease, disorder, or condition, i.e., arresting itsdevelopment; and

(iii) relieving the disease, disorder, or condition, i.e., causingregression of the disease, disorder, and/or condition.

The terms “R” and “S” indicate the specific stereochemical configurationof a substituent at an asymmetric carbon atom in a chemical structure asdrawn.

The term “rac” indicates that a compound is a racemate, which is definedas an equimolar mixture of a pair of enantiomers. A “rac” compound doesnot exhibit optical activity. The chemical name or formula of a racemateis distinguished from those of the enantiomers by the prefix (±)- orrac- (or racem-) or by the symbols RS and SR.

The terms “endo” and “exo” are descriptors of the relative orientationof substituents attached to non-bridgehead atoms in abicyclo[x.y.z]alkane (x≧y>z>0).

The terms “syn” and “anti” are descriptors of the relative orientationof substituents attached to bridgehead atoms in a bicyclo[x.y.z]alkane(x≧y>z>0).

The term “exo” is given to a substituent (e.g., Br attached to C-2 inthe example below) that is orientated towards the highest numberedbridge (z bridge, e.g., C-7 in example below); if the substituent isorientated away from the highest numbered bridge it is given thedescription “endo”.

The term “syn” is given to a substituent attached to the highestnumbered bridge (z bridge, e.g., F attached to C-7 in the example below)and is orientated towards the lowest numbered bridge (x bridge, e.g.,C-2 and C-3 in example below); if the substiuent is orientated away fromthe lowest numbered bridge it is given the description “anti.”

The terms “cis” and “trans” are descriptors which show the relationshipbetween two ligands attached to separate atoms that are connected by adouble bond or are contained in a ring. The two ligands are said to belocated cis to each other if they lie on the same side of a plane. Ifthey are on opposite sides, their relative position is described astrans. The appropriate reference plane of a double bond is perpendicularto that of the relevant σ-bonds and passes through the double bond. Fora ring it is the mean plane of the ring(s).

The compounds of the invention may exhibit the phenomenon oftautomerism. While Formula I cannot expressly depict all possibletautomeric forms, it is to be understood that Formula I is intended torepresent any tautomeric form of the depicted compound and is not to belimited merely to a specific compound form depicted by the formuladrawings. For illustration, and in no way limiting the range oftautomers, the compounds of Formula I may exist as the following:

Some of the inventive compounds may exist as single stereoisomers (i.e.,essentially free of other stereoisomers), racemates, and/or mixtures ofenantiomers and/or diastereomers. All such single stereoisomers,racemates and mixtures thereof are intended to be within the scope ofthe present invention. Preferably, the inventive compounds that areoptically active are used in optically pure form.

As generally understood by those skilled in the art, an optically purecompound having one chiral center (i.e., one asymmetric carbon atom) isone that consists essentially of one of the two possible enantiomers(i.e., is enantiomerically pure), and an optically pure compound havingmore than one chiral center is one that is both diastereomerically pureand enantiomerically pure. Preferably, the compounds of the presentinvention are used in a form that is at least 90% free of otherenantiomers or diastereomers of the compounds, that is, a form thatcontains at least 90% of a single isomer (80% enantiomeric excess(“e.e.”) or diastereomeric excess (“d.e.”)), more preferably at least95% (90% e.e. or d.e.), even more preferably at least 97.5% (95% e.e. ord.e.), and most preferably at least 99% (98% e.e. or d.e.).

Additionally, the Formula I is intended to cover solvated as well asunsolvated forms of the identified structures. For example, Formula Iincludes compounds of the indicated structure in both hydrated andnon-hydrsated forms. Other examples of solvates include the structuresin combination with isopropanol, ethanol, methanol, DMSO, ethyl acetate,pentyl acetate, acetic acid, or ethanolamine.

In addition to compounds of Formula I, the invention includespharmaceutically acceptable prodrugs, pharmaceutically activemetabolites, and pharmaceutically acceptable salts of such compounds andmetabolites.

“A pharmaceutically acceptable prodrug” is a compound that may beconverted under physiological conditions or by solvolysis to thespecified compound or to a pharmaceutically acceptable salt of suchcompound prior to exhibiting its pharmacological effect (s). Typically,the prodrug is formulated with the objective(s) of improved chemicalstability, improved patient acceptance and compliance, improvedbioavailability, prolonged duration of action, improved organselectivity, improved formulation (e.g., increased hydrosolubility),and/or decreased side effects (e.g., toxicity). The prodrug can bereadily prepared from the compounds of Formula I using methods known inthe art, such as those described by Burger's Medicinal Chemistry andDrug Chemistry, 1, 172-178, 949-982 (1995). See also Bertolini et al.,J. Med. Chem., 40, 2011-2016 (1997); Shan, et al., J. Pharm. Sci., 86(7), 765-767; Bagshawe, Drug Dev. Res., 34, 220-230 (1995); Bodor,Advances in Drug Res., 13, 224-331 (1984); Bundgaard, Design of Prodrugs(Elsevier Press 1985); Larsen, Design and Application of Prodrugs, DrugDesign and Development (Krogsgaard-Larsen et al., eds., Harwood AcademicPublishers, 1991); Dear et al., J. Chromatogr. B, 748, 281-293 (2000);Spraul et al., J. Pharmaceutical & Biomedical Analysis, 10, 601-605(1992); and Prox et al., Xenobiol., 3, 103-112 (1992).

“A pharmaceutically active metabolite” is intended to mean apharmacologically active product produced through metabolism in the bodyof a specified compound or salt thereof. After entry into the body, mostdrugs are substrates for chemical reactions that may change theirphysical properties and biologic effects. These metabolic conversions,which usually affect the polarity of the Formula I compounds, alter theway in which drugs are distributed in and excreted from the body.However, in some cases, metabolism of a drug is required for therapeuticeffect. For example, anticancer drugs of the anti-metabolite class mustbe converted to their active forms after they have been transported intoa cancer cell.

Since most drugs undergo metabolic transformation of some kind, thebiochemical reactions that play a role in drug metabolism may benumerous and diverse. The main site of drug metabolism is the liver,although other tissues may also participate.

A feature characteristic of many of these transformations is that themetabolic products, or “metabolites,” are more polar than the parentdrugs, although a polar drug does sometime yield a less polar product.Substances with high lipid/water partition coefficients, which passeasily across membranes, also diffuse back readily from tubular urinethrough the renal tubular cells into the plasma. Thus, such substancestend to have a low renal clearance and a long persistence in the body.If a drug is metabolized to a more polar compound, one with a lowerpartition coefficient, its tubular reabsorption will be greatly reduced.Moreover, the specific secretory mechanisms for anions and cations inthe proximal renal tubules and in the parenchymal liver cells operateupon highly polar substances.

As a specific example, phenacetin (acetophenetidin) and acetanilide areboth mild analgesic and antipyretic agents, but are transformed withinthe body to a more polar and more effective metabolite,p-hydroxyacetanilid (acetaminophen), which is widely used today. When adose of acetanilide is given to a person, the successive metabolitespeak and decay in the plasma sequentially. During the first hour,acetanilide is the principal plasma component. In the second hour, asthe acetanilide level falls, the metabolite acetaminophen concentrationreaches a peak. Finally, after a few hours, the principal plasmacomponent is a further metabolite that is inert and can be excreted fromthe body. Thus, the plasma concentrations of one or more metabolites, aswell as the drug itself, can be pharmacologically important.

“A pharmaceutically acceptable salt” is intended to mean a salt thatretains the biological effectiveness of the free acids and bases of thespecified compound and that is not biologically or otherwiseundesirable. A compound of the invention may possess a sufficientlyacidic, a sufficiently basic, or both functional groups, and accordinglyreact with any of a number of inorganic or organic bases, and inorganicand organic acids, to form a pharmaceutically acceptable salt. Exemplarypharmaceutically acceptable salts include those salts prepared byreaction of the compounds of the present invention with a mineral ororganic acid or an inorganic base, such as salts including sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates,citrates, lactates, γ-hydroxybutyrates, glycolates, tartrates,methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, and mandelates.

If the inventive compound is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, for example, treatment of the free base with an inorganic acid,such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like, or with an organic acid, such as aceticacid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonicacid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, apyranosidyl acid, such as glucuronic acid or galacturonic acid, anα-hydroxy acid, such as citric acid or tartaric acid, an amino acid,such as aspartic acid or glutamic acid, an aromatic acid, such asbenzoic acid or cinnamic acid, a sulfonic acid, such asp-toluenesulfonic acid or ethanesulfonic acid, or the like.

If the inventive compound is an acid, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method, for example,treatment of the free acid with an inorganic or organic base, such as anamine (primary, secondary or tertiary), an alkali metal hydroxide oralkaline earth metal hydroxide, or the like. Illustrative examples ofsuitable salts include organic salts derived from amino acids, such asglycine and arginine, ammonia, primary, secondary, and tertiary amines,and cyclic amines, such as piperidine, morpholine and piperazine, andinorganic salts derived from sodium, calcium, potassium, magnesium,manganese, iron, copper, zinc, aluminum and lithium.

In the case of agents that are solids, it is understood by those skilledin the art that the inventive compounds and salts may exist in differentcrystal, co-crystal, or polymorphic forms, all of which are intended tobe within the scope of the present invention and specified formulas.

Methods of Treatment and Prevention of Hepatitis C Viral Infections

The present invention provides methods for treating or preventing ahepatitis C virus infection in a patient in need thereof.

The present invention further provides methods for introducing atherapeutically effective amount of the Formula I compound orcombination of such compounds into the blood stream of a patient in thetreatment and/or prevention of hepatitis C viral infections.

The magnitude of a prophylactic or therapeutic dose of a Formula Icompound of the invention or a pharmaceutically acceptable salt,solvate, or hydrate, thereof in the acute or chronic treatment orprevention of an infection will vary, however, with the nature andseverity of the infection, and the route by which the active ingredientis administered. The dose, and in some cases the dose frequency, willalso vary according to the infection to be treated, the age, bodyweight, and response of the individual patient. Suitable dosing regimenscan be readily selected by those skilled in the art with dueconsideration of such factors.

The methods of the present invention are particularly well suited forhuman patients. In particular, the methods and doses of the presentinvention can be useful for immunocompromised patients including, butnot limited to cancer patients, HIV infected patients, and patients withan immunodegenerative disease. Furthermore, the methods can be usefulfor immunocompromised patients currently in a state of remission. Themethods and doses of the present invention are also useful for patientsundergoing other antiviral treatments. The prevention methods of thepresent invention are particularly useful for patients at risk of viralinfection. These patients include, but are not limited to health careworkers, e.g., doctors, nurses, hospice care givers; military personnel;teachers; childcare workers; patients traveling to, or living in,foreign locales, in particular third world locales including social aidworkers, missionaries, and foreign diplomats. Finally, the methods andcompositions include the treatment of refractory patients or patientsresistant to treatment such as resistance to reverse transcriptaseinhibitors, protease inhibitors, etc.

Doses

Toxicity and efficacy of the compounds of the invention can bedetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, e.g., for determining the LD₅₀ (the dose lethal to50% of the population) and the ED₅₀ (the dose therapeutically effectivein 50% of the population). The dose ratio between toxic and therapeuticeffects is the therapeutic index and it can be expressed as the ratioLD₅₀/ED₅₀.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage of the compounds for use inhumans. The dosage of such compounds lies preferably within a range ofcirculating concentrations that include the ED₅₀ with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized. For anycompound used in the method of the invention, the therapeuticallyeffective dose can be estimated initially from cell culture assays. Adose may be formulated in animal models to achieve a circulating plasmaconcentration range that includes the IC₅₀ (i.e., the concentration ofthe test compound that achieves a half-maximal inhibition of symptoms)as determined in cell culture; alternatively, the dose of the Formula Icompound may be formulated in animal models to achieve a circulatingplasma concentration range of the compound that corresponds to theconcentration required to achieve a fixed magnitude of response. Suchinformation can be used to more accurately determine useful doses inhumans. Levels in plasma may be measured, for example, by highperformance liquid chromatography.

The protocols and compositions of the invention are preferably tested invitro, and then in vivo, for the desired therapeutic or prophylacticactivity, prior to use in humans. For example, in vitro assays which canbe used to determine whether administration of a specific therapeuticprotocol is indicated, include in vitro cell culture assays in whichcells that are responsive to the effects of the Formula I compounds areexposed to the ligand and the magnitude of response is measured by anappropriate technique. The assessment of the Formula I compound is thenevaluated with respect to the Formula I compound potency, and the degreeof conversion of the Formula I compound prodrug. Compounds for use inmethods of the invention can be tested in suitable animal model systemsprior to testing in humans, including but not limited to in rats, mice,chicken, cows, monkeys, rabbits, hamsters, etc. The compounds can thenbe used in the appropriate clinical trials.

The magnitude of a prophylactic or therapeutic dose of a prodrug of aFormula I compound of the invention or a pharmaceutically acceptablesalt, solvate, or hydrate thereof in the acute or chronic treatment orprevention of an infection or condition will vary with the nature andseverity of the infection, and the route by which the active ingredientis administered. The dose, and perhaps the dose frequency, will alsovary according to the infection to be treated, the age, body weight, andresponse of the individual patient. Suitable dosing regimens can bereadily selected by those skilled in the art with due consideration ofsuch factors. In one embodiment, the dose administered depends upon thespecific compound to be used, and the weight and condition of thepatient. Also, the dose may differ for various particular Formula Icompounds; suitable doses can be predicted on the basis of theaforementioned in vitro measurements and on the basis of animal studies,such that smaller doses will be suitable for those Formula I compoundsthat show effectiveness at lower concentrations than other Formula Icompounds when measured in the systems described or referenced herein.In general, the dose per day is in the range of from about 0.001 to 100mg/kg, preferably about 1 to 25 mg/kg, more preferably about 5 to 15mg/kg. For treatment of humans infected by hepatitis C viruses, about0.1 mg to about 15 g per day is administered in about one to fourdivisions a day, preferably 100 mg to 12 g per day, more preferably from100 mg to 8000 mg per day.

Additionally, the recommended daily dose ran can be administered incycles as single agents or in combination with other therapeutic agents.In one embodiment, the daily dose is administered in a single dose or inequally divided doses. In a related embodiment, the recommended dailydose can be administered once time per week, two times per week, threetimes per week, four times per week or five times per week.

In one embodiment, the compounds of the invention are administered toprovide systemic distribution of the compound within the patient. In arelated embodiment, the compounds of the invention are administered toproduce a systemic effect in the body.

In another embodiment the compounds of the invention are administeredvia oral, mucosal (including sublingual, buccal, rectal, nasal, orvaginal), parenteral (including subcutaneous, intramuscular, bolusinjection, intraarterial, or intravenous), transdermal, or topicaladministration. In a specific embodiment the compounds of the inventionare administered via mucosal (including sublingual, buccal, rectal,nasal, or vaginal), parenteral (including subcutaneous, intramuscular,bolus injection, intraarterial, or intravenous), transdermal, or topicaladministration. In a further specific embodiment, the compounds of theinvention are administered via oral administration. In a furtherspecific embodiment, the compounds of the invention are not administeredvia oral administration.

Different therapeutically effective amounts may be applicable fordifferent infections, as will be readily known by those of ordinaryskill in the art. Similarly, amounts sufficient to treat or prevent suchinfections, but insufficient to cause, or sufficient to reduce, adverseeffects associated with conventional therapies are also encompassed bythe above described dosage amounts and dose frequency schedules.

Combination Therapy

Specific methods of the invention further comprise the administration ofan additional therapeutic agent (i.e., a therapeutic agent other than acompound of the invention). In certain embodiments of the presentinvention, the compounds of the invention can be used in combinationwith at least one other therapeutic agent. Therapeutic agents include,but are not limited to antibiotics, antiemetic agents, antidepressants,and antifungal agents, anti-inflammatory agents, antiviral agents,anticancer agents, immunomodulatory agents, α-interferons,β-interferons, ribavirin, alkylating agents, hormones, cytokines, ortoll-like receptor modulators. In one embodiment the inventionencompasses the administration of an additional therapeutic agent thatis HCV specific or demonstrates anti-HCV activity.

The Formula I compounds of the invention can be administered orformulated in combination with antibiotics. For example, they can beformulated with a macrolide (e.g., tobramycin (Tobi®)), a cephalosporin(e.g., cephalexin (Keflex®), cephradine (Velosef®), cefuroxime(Ceftin®), cefprozil (Cefzil®), cefaclor (Ceclor®), cefixime (Suprax®)or cefadroxil (Duricef®)), a clarithromycin (e.g., clarithromycin(Biaxin®)), an erythromycin (e.g., erythromycin (EMycin®)), a penicillin(e.g., penicillin V (V-Cillin K® or Pen Vee K®)) or a quinolone (e.g.,ofloxacin (Floxin®), ciprofloxacin (Cipro®) or norfloxacin (Noroxin®)),aminoglycoside antibiotics (e.g., apramycin, arbekacin, bambermycins,butirosin, dibekacin, neomycin, neomycin, undecylenate, netilmicin,paromomycin, ribostamycin, sisomicin, and spectinomycin), amphenicolantibiotics (e.g., azidamfenicol, chloramphenicol, florfenicol, andthiamphenicol), ansamycin antibiotics (e.g., rifamide and rifampin),carbacephems (e.g., loracarbef), carbapenems (e.g., biapenem andimipenem), cephalosporins (e.g., cefaclor, cefadroxil, cefamandole,cefatrizine, cefazedone, cefozopran, cefpimizole, cefpiramide, andcefpirome), cephamycins (e.g., cefbuperazone, cefmetazole, andcefminox), monobactams (e.g., aztreonam, carumonam, and tigemonam),oxacephems (e.g., flomoxef, and moxalactam), penicillins (e.g.,amdinocillin, amdinocillin pivoxil, amoxicillin, bacampicillin,benzylpenicillinic acid, benzylpenicillin sodium, epicillin,fenbenicillin, floxacillin, penamccillin, penethamate hydriodide,penicillin o-benethamine, penicillin 0, penicillin V, penicillin Vbenzathine, penicillin V hydrabamine, penimepicycline, andphencihicillin potassium), lincosamides (e.g., clindamycin, andlincomycin), amphomycin, bacitracin, capreomycin, colistin, enduracidin,enviomycin, tetracyclines (e.g., apicycline, chlortetracycline,clomocycline, and demeclocycline), 2,4-diaminopyrimidines (e.g.,brodimoprim), nitrofurans (e.g., furaltadone, and furazolium chloride),quinolones and analogs thereof (e.g., cinoxacin, clinafloxacin,flumequine, and grepagloxacin), sulfonamides (e.g., acetylsulfamethoxypyrazine, benzylsulfamide, noprylsulfamide,phthalylsulfacetamide, sulfachrysoidine, and sulfacytine), sulfones(e.g., diathymosulfone, glucosulfone sodium, and solasulfone),cycloserine, mupirocin and tuberin.

The Formula I compounds of the invention can also be administered orformulated in combination with an antiemetic agent. Suitable antiemeticagents include, but are not limited to, metoclopromide, domperidone,prochlorperazine, promethazine, chlorpromazine, trimethobenzamide,ondansetron, granisetron, hydroxyzine, acetylleucine monoethanolamine,alizapride, azasetron, benzquinamide, bietanautine, bromopride,buclizine, clebopride, cyclizine, dimenhydrinate, diphenidol,dolasetron, meclizine, methallatal, metopimazine, nabilone, oxyperndyl,pipamazine, scopolamine, sulpiride, tetrahydrocannabinols,thiethylperazine, thioproperazine, tropisetron, and mixtures thereof.

The Formula I compounds of the invention can be administered orformulated in combination with an antidepressant. Suitableantidepressants include, but are not limited to, binedaline, caroxazone,citalopram, dimethazan, fencamine, indalpine, indeloxazinehydrochloride, nefopam, nomifensine, oxitriptan, oxypertine, paroxetine,sertraline, thiazesim, trazodone, benmoxine, iproclozide, iproniazid,isocarboxazid, nialamide, octamoxin, phenelzine, cotinine, rolicyprine,rolipram, maprotiline, metralindole, mianserin, mirtazepine, adinazolam,amitriptyline, amitriptylinoxide, amoxapine, butriptyline, clomipramine,demexiptiline, desipramine, dibenzepin, dimetacrine, dothiepin, doxepin,fluacizine, imipramine, imipramine N-oxide, iprindole, lofepramine,melitracen, metapramine, nortriptyline, noxiptilin, opipramol,pizotyline, propizepine, protriptyline, quinupramine, tianeptine,trimipramine, adrafinil, benactyzine, bupropion, butacetin, dioxadrol,duloxetine, etoperidone, febarbamate, femoxetine, fenpentadiol,fluoxetine, fluvoxamine, hematoporphyrin, hypericin, levophacetoperane,medifoxamine, milnacipran, minaprine, moclobemide, nefazodone,oxaflozane, piberaline, prolintane, pyrisuccideanol, ritanserin,roxindole, rubidium chloride, sulpiride, tandospirone, thozalinone,tofenacin, toloxatone, tranylcypromine, L-tryptophan, venlafaxine,viloxazine, and zimeldine.

The Formula I compounds of the invention can be administered orformulated in combination with an antifungal agent. Suitable antifungalagents include but are not limited to amphotericin B, itraconazole,ketoconazole, fluconazole, intrathecal, flucytosine, miconazole,butoconazole, clotrimazole, nystatin, terconazole, tioconazole,ciclopirox, econazole, haloprogrin, naftifine, terbinafine,undecylenate, and griseofulvin.

The Formula I compounds of the invention can be administered orformulated in combination with an anti-inflammatory agent. Usefulanti-inflammatory agents include, but are not limited to, non-steroidalanti-inflammatory drugs such as salicylic acid, acetylsalicylic acid,methyl salicylate, diflunisal, salsalate, olsalazine, sulfasalazine,acetaminophen, indomethacin, sulindac, etodolac, mefenamic acid,meclofenamate sodium, tolmetin, ketorolac, dichlofenac, ibuprofen,naproxen, naproxen sodium, fenoprofen, ketoprofen, flurbinprofen,oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam, pivoxicam,tenoxicam, nabumetome, phenylbutazone, oxyphenbutazone, antipyrine,aminopyrine, apazone and nimesulide; leukotriene antagonists including,but not limited to, zileuton, aurothioglucose, gold sodium thiomalateand auranofin; steroids including, but not limited to, alclometasonediproprionate, amcinonide, beclomethasone dipropionate, betametasone,betamethasone benzoate, betamethasone diproprionate, betamethasonesodium phosphate, betamethasone valerate, clobetasol proprionate,clocortolone pivalate, hydrocortisone, hydrocortisone derivatives,desonide, desoximatasone, dexamethasone, flunisolide, flucoxinolide,flurandrenolide, halcinocide, medrysone, methylprednisolone,methprednisolone acetate, methylprednisolone sodium succinate,mometasone furoate, paramethasone acetate, prednisolone, prednisoloneacetate, prednisolone sodium phosphate, prednisolone tebuatate,prednisone, triamcinolone, triamcinolone acetonide, triamcinolonediacetate, and triamcinolone hexacetonide; and other anti-inflammatoryagents including, but not limited to, methotrexate, colchicine,allopurinol, probenecid, sulfinpyrazone and benzbromarone.

The Formula I compounds of the invention can be administered orformulated in combination with another antiviral agent. Useful antiviralagents include, but are not limited to, protease inhibitors, nucleosidereverse transcriptase inhibitors, non-nucleoside reverse transcriptaseinhibitors and nucleoside analogs. The antiviral agents include but arenot limited to zidovudine, acyclovir, gangcyclovir, vidarabine,idoxuridine, trifluridine, levovirin, viramidine and ribavirin, as wellas foscarnet, amantadine, rimantadine, saquinavir, indinavir,amprenavir, lopinavir, ritonavir, the α-interferons, β-interferons,adefovir, clevadine, entecavir, pleconaril.

The Formula I compound of the invention can be administered orformulated in combination with an immunomodulatory agent.Immunomodulatory agents include, but are not limited to, methothrexate,leflunomide, cyclophosphamide, cyclosporine A, mycophenolate mofetil,rapamycin (sirolimus), mizoribine, deoxyspergualin, brequinar,malononitriloamindes (e.g., leflunamide), T cell receptor modulators,and cytokine receptor modulators, peptide mimetics, and antibodies(e.g., human, humanized, chimeric, monoclonal, polyclonal, Fvs, ScFvs,Fab or F(ab)₂ fragments or epitope binding fragments), nucleic acidmolecules (e.g., antisense nucleic acid molecules and triple helices),small molecules, organic compounds, and inorganic compounds. Examples ofT cell receptor modulators include, but are not limited to, anti-T cellreceptor antibodies (e.g., anti-CD4 antibodies (e.g., cM-T412(Boehringer), IDEC-CE9.1® (IDEC and SKB), mAB 4162W94, Orthoclone andOKTcdr4a (Janssen-Cilag)), anti-CD3 antibodies (e.g., Nuvion (ProductDesign Labs), OKT3 (Johnson & Johnson), or Rituxan (IDEC)), anti-CD5antibodies (e.g., an anti-CD5 ricin-linked immunoconjugate), anti-CD7antibodies (e.g., CHH-380 (Novartis)), anti-CD8 antibodies, anti-CD40ligand monoclonal antibodies (e.g., IDEC-131 (IDEC)), anti-CD52antibodies (e.g., CAMPATH 1H (Ilex)), anti-CD2 antibodies, anti-CD11aantibodies (e.g., Xanelim (Genentech)), anti-B7 antibodies (e.g.,IDEC-114 (IDEC)), CTLA4-immunoglobulin, and toll-like receptor (TLR)modulators. Examples of cytokine receptor modulators include, but arenot limited to, soluble cytokine receptors (e.g., the extracellulardomain of a TNF-α receptor or a fragment thereof, the extracellulardomain of an IL-β receptor or a fragment thereof, and the extracellulardomain of an IL-6 receptor or a fragment thereof), cytokines orfragments thereof (e.g., interleukin (IL)-2, IL-3, IL-4, IL-5, IL-6,IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-15, TNF-α, interferon (IFN)-α,IFN-β, IFN-γ, and GM-CSF), anti-cytokine receptor antibodies (e.g.,anti-IFN receptor antibodies, anti-IL-2 receptor antibodies (e.g.,Zenapax (Protein Design Labs)), anti-IL-4 receptor antibodies, anti-IL-6receptor antibodies, anti-IL-10 receptor antibodies, and anti-IL-12receptor antibodies), anti-cytokine antibodies (e.g., anti-IFNantibodies, anti-TNF-α antibodies, anti-IL-1β antibodies, anti-IL-6antibodies, anti-IL-8 antibodies (e.g., ABX-IL-8 (Abgenix)), andanti-IL-12 antibodies).

The Formula I compounds of the invention can be administered orformulated in combination with an agent which inhibits viral enzymes,including but not limited to inhibitors of HCV protease, such as BILN2061, SCH-503034, ITMN-191, or VX-950; and inhibitors of NS5B polymerasesuch as NM107 (and its prodrug NM283), R1626, R7078, BILN1941,GSK625433, GILD9128 or HCV-796.

The Formula I compounds of the invention can be administered orformulated in combination with an agent which inhibits HCV polymerasesuch as those described in Wu, Curr Drug Targets Infect Disord. 2003,3(3), 207-19 or in combination with compounds that inhibit the helicasefunction of the virus such as those described in Bretner M, et al.Nucleosides Nucleotides Nucleic Acids. 2003, 22(5-8), 1531, or withinhibitors of other HCV specific targets such as those described inZhang X., IDrugs 2002, 5(2), 154-8.

The Formula I compounds of the invention can be administered orformulated in combination with an agent which inhibits viralreplication.

The Formula I compounds of the invention can be administered orformulated in combination with cytokines. Examples of cytokines include,but are not limited to, interleukin-2 (IL-2), interleukin-3 (IL-3),interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6),interleukin-7 (IL-7), interleukin-9 (IL-9), interleukin-10 (IL-10),interleukin-12 (IL-12), interleukin 15 (IL-15), interleukin 18 (IL-18),platelet derived growth factor (PDGF), erythropoietin (Epo), epidermalgrowth factor (EGF), fibroblast growth factor (FGF), granulocytemacrophage stimulating factor (GM-CSF), granulocyte colony stimulatingfactor (G-CSF), macrophage colony stimulating factor (M-CSF), prolactin,and interferon (IFN), e.g., IFN-α, and IFN-γ).

The Formula I compounds of the invention can be administered orformulated in combination with hormones. Examples of hormones include,but are not limited to, luteinizing hormone releasing hormone (LHRH),growth hormone (GH), growth hormone releasing hormone, ACTH,somatostatin, somatotropin, somatomedin, parathyroid hormone,hypothalamic releasing factors, insulin, glucagon, enkephalins,vasopressin, calcitonin, heparin, low molecular weight heparins,heparinoids, synthetic and natural opioids, insulin thyroid stimulatinghormones, and endorphins.

The Formula I compounds of the invention can be administered orformulated in combination with β-interferons which include, but are notlimited to, interferon β-1a, interferon β-1b.

The Formula I compounds of the invention can be administered orformulated in combination with α-interferons which include, but are notlimited to, interferon α-1, interferon α-2a (roferon), interferon α-2b,intron, Peg-Intron, Pegasys, consensus interferon (infergen) andalbuferon.

The Formula I compounds of the invention can be administered orformulated in combination with an absorption enhancer, particularlythose which target the lymphatic system, including, but not limited tosodium glycocholate; sodium caprate; N-lauryl-β-D-maltopyranoside; EDTA;mixed micelle; and those reported in Muranishi Crit. Rev. Ther. DrugCarrier Syst., 7, 1-33, which is hereby incorporated by reference in itsentirety. Other known absorption enhancers can also be used. Thus, theinvention also encompasses a pharmaceutical composition comprising oneor more Formula I compounds of the invention and one or more absorptionenhancers.

The Formula I compounds of the invention can be administered orformulated in combination with a cytochrome P450 monooxygenaseinhibitor, such as, but not limited to, ritonavir or a pharmaceuticallyacceptable salt, ester, and prodrug thereof to improve thepharmacokinetics (e.g., increased half-life, increased time to peakplasma concentration, increased blood levels) of a Formula I compoundthat is metabolized by cytochrome P450 monooxygenase. Thus, theinvention also encompasses a pharmaceutical composition comprisingFormula I compounds of the invention and one or more cytochrome P450monooxygenase inhibitors.

The Formula I compounds of the invention can be administered incombination with food to enhance absorption of the Formula I compoundsin the gastrointestinal tract and to increase the bioavailability of theFormula I compounds.

The Formula I compounds of the invention can be administered orformulated in combination with an alkylating agent. Examples ofalkylating agents include, but are not limited to nitrogen mustards,ethylenimines, methylmelamines, alkyl sulfonates, nitrosoureas,triazenes, mechlorethamine, cyclophosphamide, ifosfamide, melphalan,chlorambucil, hexamethylmelaine, thiotepa, busulfan, carmustine,streptozocin, dacarbazine and temozolomide.

The compounds of the invention and the other therapeutics agent can actadditively or, more preferably, synergistically. In one embodiment, acomposition comprising a compound of the invention is administeredconcurrently with the administration of another therapeutic agent, whichcan be part of the same composition or in a different composition fromthat comprising the compounds of the invention. In another embodiment, acompound of the invention is administered prior to or subsequent toadministration of another therapeutic agent. In a separate embodiment, acompound of the invention is administered to a patient who has notpreviously undergone or is not currently undergoing treatment withanother therapeutic agent, particularly an antiviral agent.

In one embodiment, the methods of the invention comprise theadministration of one or more Formula I compounds of the inventionwithout an additional therapeutic agent.

Pharmaceutical Compositions and Dosage Forms

Pharmaceutical compositions and single unit dosage forms comprising aFormula I compound of the invention, or a pharmaceutically acceptablesalt, or hydrate thereof, are also encompassed by the invention.Individual dosage forms of the invention may be suitable for oral,mucosal (including sublingual, buccal, rectal, nasal, or vaginal),parenteral (including subcutaneous, intramuscular, bolus injection,intraarterial, or intravenous), transdermal, or topical administration.Pharmaceutical compositions and dosage forms of the invention typicallyalso comprise one or more pharmaceutically acceptable excipients.Sterile dosage forms are also contemplated.

In an alternative embodiment, pharmaceutical composition encompassed bythis embodiment includes a Formula I compound of the invention, or apharmaceutically acceptable salt, or hydrate thereof, and at least oneadditional therapeutic agent. Examples of additional therapeutic agentsinclude, but are not limited to, those listed above.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage form usedin the acute treatment of a disease or a related disease may containlarger amounts of one or more of the active ingredients it comprisesthan a dosage form used in the chronic treatment of the same disease.Similarly, a parenteral dosage form may contain smaller amounts of oneor more of the active ingredients it comprises than an oral dosage formused to treat the same disease or disorder. These and other ways inwhich specific dosage forms encompassed by this invention will vary fromone another will be readily apparent to those skilled in the art. See,e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing,Easton Pa. (1990). Examples of dosage forms include, but are not limitedto: tablets; caplets; capsules, such as soft elastic gelatin capsules;cachets; troches; lozenges; dispersions; suppositories; ointments;cataplasms (poultices); pastes; powders; dressings; creams; plasters;solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels;liquid dosage forms suitable for oral or mucosal administration to apatient, including suspensions (e.g., aqueous or non-aqueous liquidsuspensions, oil-in-water emulsions, or a water-in-oil liquidemulsions), solutions, and elixirs; liquid dosage forms suitable forparenteral administration to a patient; and sterile solids (e.g.,crystalline or amorphous solids) that can be reconstituted to provideliquid dosage forms suitable for parenteral administration to a patient.

Typical pharmaceutical compositions and dosage forms comprise one ormore carriers, excipients or diluents. Suitable excipients are wellknown to those skilled in the art of pharmacy, and non-limiting examplesof suitable excipients are provided herein. Whether a particularexcipient is suitable for incorporation into a pharmaceuticalcomposition or dosage form depends on a variety of factors well known inthe art including, but not limited to, the way in which the dosage formwill be administered to a patient. For example, oral dosage forms suchas tablets may contain excipients not suited for use in parenteraldosage forms. The suitability of a particular excipient may also dependon the specific active ingredients in the dosage form.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Carstensen, Drug Stability: Principles & Practice, 2d.Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect, water andheat accelerate the decomposition of some compounds. Thus, the effect ofwater on a formulation can be of great significance since moistureand/or humidity are commonly encountered during manufacture, handling,packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizers,” include, but are not limited to,antioxidants such as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms of the invention compriseFormula I compounds of the invention, or a pharmaceutically acceptablesalt or hydrate thereof comprise 0.1 mg to 1500 mg per unit to providedoses of about 0.01 to 200 mg/kg per day.

Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an intimate admixture with at least oneexcipient according to conventional pharmaceutical compoundingtechniques. Excipients can take a wide variety of forms depending on theform of preparation desired for administration. For example, excipientssuitable for use in oral liquid or aerosol dosage forms include, but arenot limited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, PA), and mixtures thereof. Aspecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103™ and Starch 1500LM.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,specifically from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, pre-gelatinized starch, otherstarches, clays, other algins, other celluloses, gums, and mixturesthereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

Delayed Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry and/or lyophylized products ready tobe dissolved or suspended in a pharmaceutically acceptable vehicle forinjection (reconstitutable powders), suspensions ready for injection,and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

Transdermal Dosage Forms

Transdermal dosage forms include “reservoir type” or “matrix type”patches, which can be applied to the skin and worn for a specific periodof time to permit the penetration of a desired amount of activeingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal and topical dosage formsencompassed by this invention are well known to those skilled in thepharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

Topical Dosage Forms

Topical dosage forms of the invention include, but are not limited to,creams, lotions, ointments, gels, solutions, emulsions, suspensions, orother forms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences, 18th eds., Mack Publishing, Easton Pa. (1990);and Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger,Philadelphia (1985).

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal and topical dosage formsencompassed by this invention are well known to those skilled in thepharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

Mucosal Dosage Forms

Mucosal dosage forms of the invention include, but are not limited to,ophthalmic solutions, sprays and aerosols, or other forms known to oneof skill in the art. See, e.g., Remington's Pharmaceutical Sciences,18th eds., Mack Publishing, Easton Pa. (1990); and Introduction toPharmaceutical Dosage Forms, 4th ed., Lea & Febiger, Philadelphia(1985). Dosage forms suitable for treating mucosal tissues within theoral cavity can be formulated as mouthwashes or as oral gels. In oneembodiment, the aerosol comprises a carrier. In another embodiment, theaerosol is carrier free.

The Formula I compounds of the invention may also be administereddirectly to the lung by inhalation. For administration by inhalation, aFormula I compound can be conveniently delivered to the lung by a numberof different devices. For example, a Metered Dose Inhaler (“MDI”) whichutilizes canisters that contain a suitable low boiling propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas can beused to deliver a Formula I compound directly to the lung. MDI devicesare available from a number of suppliers such as 3M Corporation,Aventis, Boehringer Ingelheim, Forest Laboratories, Glaxo-Wellcome,Schering Plough and Vectura.

Alternatively, a Dry Powder Inhaler (DPI) device can be used toadminister a Formula I compound to the lung (see, e.g., Raleigh et al.,Proc. Amer. Assoc. Cancer Research Annual Meeting, 1999, 40, 397, whichis herein incorporated by reference). DPI devices typically use amechanism such as a burst of gas to create a cloud of dry powder insidea container, which can then be inhaled by the patient. DPI devices arealso well known in the art and can be purchased from a number of vendorswhich include, for example, Fisons, Glaxo-Wellcome, Inhale TherapeuticSystems, ML Laboratories, Qdose and Vectura. A popular variation is themultiple dose DPI (“MDDPI”) system, which allows for the delivery ofmore than one therapeutic dose. MDDPI devices are available fromcompanies such as AstraZeneca, GlaxoWellcome, IVAX, Schering Plough,SkyePharma and Vectura. For example, capsules and cartridges of gelatinfor use in an inhaler or insufflator can be formulated containing apowder mix of the compound and a suitable powder base such as lactose orstarch for these systems.

Another type of device that can be used to deliver a Formula I compoundto the lung is a liquid spray device supplied, for example, by AradigmCorporation. Liquid spray systems use extremely small nozzle holes toaerosolize liquid drug formulations that can then be directly inhaledinto the lung.

In one embodiment, a nebulizer device is used to deliver a Formula Icompound to the lung. Nebulizers create aerosols from liquid drugformulations by using, for example, ultrasonic energy to form fineparticles that can be readily inhaled (See e.g., Verschoyle et al.,British J. Cancer, 1999, 80, Suppl 2, 96, which is herein incorporatedby reference). Examples of nebulizers include devices supplied bySheffield/Systemic Pulmonary Delivery Ltd. (See, Armer et al., U.S. Pat.No. 5,954,047; van der Linden et al., U.S. Pat. No. 5,950,619; van derLinden et al., U.S. Pat. No. 5,970,974, which are herein incorporated byreference), Aventis and Batelle Pulmonary Therapeutics.

In one embodiment, an electrohydrodynamic (“EHD”) aerosol device is usedto deliver Formula I compounds to the lung. EHD aerosol devices useelectrical energy to aerosolize liquid drug solutions or suspensions(see, e.g., Noakes et al., U.S. Pat. No. 4,765,539; Coffee, U.S. Pat.No., 4,962,885; Coffee, PCT Application, WO 94/12285; Coffee, PCTApplication, WO 94/14543; Coffee, PCT Application, WO 95/26234, Coffee,PCT Application, WO 95/26235, Coffee, PCT Application, WO 95/32807,which are herein incorporated by reference). The electrochemicalproperties of the Formula I compounds formulation may be importantparameters to optimize when delivering this drug to the lung with an EHDaerosol device and such optimization is routinely performed by one ofskill in the art. EHD aerosol devices may more efficiently deliverydrugs to the lung than existing pulmonary delivery technologies. Othermethods of intra-pulmonary delivery of Formula I compounds will be knownto the skilled artisan and are within the scope of the invention.

Liquid drug formulations suitable for use with nebulizers and liquidspray devices and EHD aerosol devices will typically include a Formula Icompound with a pharmaceutically acceptable carrier. Preferably, thepharmaceutically acceptable carrier is a liquid such as alcohol, water,polyethylene glycol or a perfluorocarbon. Optionally, another materialmay be added to alter the aerosol properties of the solution orsuspension of the Formula I compound. Preferably, this material isliquid such as an alcohol, glycol, polyglycol or a fatty acid. Othermethods of formulating liquid drug solutions or suspension suitable foruse in aerosol devices are known to those of skill in the art (see,e.g., Biesalski, U.S. Pat. No. 5,112,598; Biesalski, U.S. Pat. No.5,556,611, which are herein incorporated by reference) A Formula Icompound can also be formulated in rectal or vaginal compositions suchas suppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, a Formula Icompound can also be formulated as a depot preparation. Such long actingformulations can be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, the compounds can be formulated with suitable polymeric orhydrophobic materials (for example, as an emulsion in an acceptable oil)or ion exchange resins, or as sparingly soluble derivatives, forexample, as a sparingly soluble salt.

Alternatively, other pharmaceutical delivery systems can be employed.Liposomes and emulsions are well known examples of delivery vehiclesthat can be used to deliver Formula I compounds. Certain organicsolvents such as dimethylsulfoxide can also be employed, althoughusually at the cost of greater toxicity. A Formula I compound can alsobe delivered in a controlled release system. In one embodiment, a pumpcan be used (Sefton, CRC Crit. Ref Biomed Eng., 1987, 14, 201; Buchwaldet al., Surgery, 1980, 88, 507; Saudek et al., N. Engl. J. Med., 1989,321, 574). In another embodiment, polymeric materials can be used (seeMedical Applications of Controlled Release, Langer and Wise (eds.), CRCPres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, DrugProduct Design and Performance, Smolen and Ball (eds.), Wiley, New York(1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem., 1983,23, 61; see also Levy et al., Science, 1985, 228, 190; During et al.,Ann. Neurol., 1989, 25, 351; Howard et al., J. Neurosurg., 71, 105(1989). In yet another embodiment, a controlled-release system can beplaced in proximity of the target of the compounds of the invention,e.g., the lung, thus requiring only a fraction of the systemic dose(see, e.g., Goodson, in Medical Applications of Controlled Release,supra, vol. 2, pp. 115 (1984)). Other controlled-release system can beused (see, e.g., Langer, Science, 1990, 249, 1527).

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide mucosal dosage forms encompassed by thisinvention are well known to those skilled in the pharmaceutical arts,and depend on the particular site or method which a given pharmaceuticalcomposition or dosage form will be administered. With that fact in mind,typical excipients include, but are not limited to, water, ethanol,ethylene glycol, propylene glycol, butane-1,3-diol, isopropyl myristate,isopropyl palmitate, mineral oil, and mixtures thereof, which arenon-toxic and pharmaceutically acceptable. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences, 18th eds., Mack Publishing, Easton Pa. (1990).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, canalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

Kits

The invention provides a pharmaceutical pack or kit comprising one ormore containers comprising a Formula I compound useful for the treatmentor prevention of a Hepatitis C virus infection. In other embodiments,the invention provides a pharmaceutical pack or kit comprising one ormore containers comprising a Formula I compound useful for the treatmentor prevention of a Hepatitis C virus infection and one or morecontainers comprising an additional therapeutic agent, including but notlimited to those listed above, in particular an antiviral agent, aninterferon, an agent which inhibits viral enzymes, or an agent whichinhibits viral replication, preferably the additional therapeutic agentis HCV specific or demonstrates anti-HCV activity.

The invention also provides a pharmaceutical pack or kit comprising oneor more containers comprising one or more of the ingredients of thepharmaceutical compositions of the invention. Optionally associated withsuch container(s) can be a notice in the form prescribed by agovernmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, which notice reflects approvalby the agency of manufacture, use or sale for human administration.

The inventive agents may be prepared using the reaction routes andsynthesis schemes as described below, employing the general techniquesknown in the art using starting materials that are readily available.The synthesis of non-exemplified compounds according to the inventionmay be successfully performed by modifications apparent to those skilledin the art, e.g., by appropriately protecting interfering groups, bychanging to other suitable reagents known in the art, or by makingroutine modifications of reaction conditions. Alternatively, otherreactions disclosed herein or generally known in the art will berecognized as having applicability for preparing other compounds of theinvention.

Preparation of Compounds

In the synthetic schemes described below, unless otherwise indicated alltemperatures are set forth in degrees Celsius and all parts andpercentages are by weight.

Reagents were purchased from commercial suppliers such as AldrichChemical Company or Lancaster Synthesis Ltd. and were used withoutfurther purification unless otherwise indicated. All solvents werepurchased from commercial suppliers such as Aldrich, EMD Chemicals orFisher and used as received.

The reactions set forth below were done generally under a positivepressure of argon or nitrogen at an ambient temperature (unlessotherwise stated) in anhydrous solvents, and the reaction flasks werefitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried.

The reactions were assayed by TLC and/or analyzed by LC-MS or HPLC andterminated as judged by the consumption of starting material. Analyticalthin layer chromatography (TLC) was performed on glass-plates precoatedwith silica gel 60 F₂₅₄ 0.25 mm plates (EMD Chemicals), and visualizedwith UV light (254 nm) and/or iodine on silica gel and/or heating withTLC stains such as ethanolic phosphomolybdic acid, ninhydrin solution,potassium permanganate solution or ceric sulfate solution. Preparativethin layer chromatography (prepTLC) was performed on glass-platesprecoated with silica gel 60 F₂₅₄ 0.5 mm plates (20×20 cm, from ThomsonInstrument Company) and visualized with UV light (254 nm).

Work-ups were typically done by doubling the reaction volume with thereaction solvent or extraction solvent and then washing with theindicated aqueous solutions using 25% by volume of the extraction volumeunless otherwise indicated. Product solutions were dried over anhydrousNa₂SO₄ and/or MgSO₄ prior to filtration and evaporation of the solventsunder reduced pressure on a rotary evaporator and noted as solventsremoved in vacuo. Column chromatography was completed under positivepressure using Merck silica gel 60, 230-400 mesh or 50-200 mesh neutralalumina, ISCO Flash-chromatography using prepacked RediSep silica gelcolumns, or Analogix flash column chromatography using prepackedSuperFlash silica gel columns. Hydrogenolysis was done at the pressureindicated in the examples or at ambient pressure.

¹H-NMR spectra and ¹³C-NMR were recorded on a Varian Mercury-VX400instrument operating at 400 MHz. NMR spectra were obtained as CDCl₃solutions (reported in ppm), using chloroform as the reference standard(7.27 ppm for the proton and 77.00 ppm for carbon), CD₃OD (3.4 and 4.8ppm for the protons and 49.3 ppm for carbon), DMSO-d₆ (2.49 ppm forproton), or internally tetramethylsilane (0.00 ppm) when appropriate.Other NMR solvents were used as needed. When peak multiplicities arereported, the following abbreviations are used: s (singlet), d(doublet), t (triplet), q (quartet), m (multiplet), br (broadened), bs(broad singlet), dd (doublet of doublets), dt (doublet of triplets).Coupling constants, when given, are reported in Hertz (Hz).

Infrared (IR) spectra were recorded on an ATR FT-IR Spectrometer as neatoils or solids, and when given are reported in wave numbers (cm⁻¹). Massspectra reported are (+)-ES or APCI (+) LC/MS conducted by theAnalytical Chemistry Department of Anadys Pharmaceuticals, Inc.Elemental analyses were conducted by the Atlantic Microlab, Inc. inNorcross, Ga. Melting points (mp) were determined on an open capillaryapparatus, and are uncorrected.

Enantiomeric excess (ee) values were determined by HPLC-analysis usingthe Chiralpak (Chiral Technologies Inc.) columns AS-RH, 2.1×150 mm, 5micron, λ=312 nm or AS-RH, 4.6×250 mm, 5 micron, λ=310 nm.

AS-RH, 2.1×150 mm, 5 micron: Binary gradient HPLC separation. Solvent A:0.1% Formic Acid in Water, Solvent B: 0.1% Formic Acid in Acetonitrile.Injected 10 μL of sample dissolved in 50% methanol—50% water [0.1mg/mL].

Time (min) % B Flow (mL/min) 0.0 55 0.3 5.0 95 0.3 5.5 95 0.3 6.0 55 0.312.0 55 0.3

AS-RH, 4.6×250 mm, 5 micron: Binary gradient HPLC separation. Solvent A:0.05% TFA in Water, Solvent B: 0.05% TFA in Acetonitrile. Injected 3-5μl of sample dissolved in acetonitrile [1 mg/mL].

Time (min) % B Flow (mL/min) 0.0 50 0.8 8.0 95 0.8 10.0 95 0.8 11.0 500.8 13.0 50 0.8

The described synthetic pathways and experimental procedures utilizemany common chemical abbreviations, 2,2-DMP (2,2-dimethoxypropane), Ac(acetyl), ACN (acetonitrile), Bn (benzyl), BnOH (benzyl alcohol), Boc(tert-butoxycarbonyl), Boc₂O (di-tert-butyl dicarbonate), Bz (benzoyl),CSI (chlorosulfonyl isocyanate), DBU(1,8-diazabicyclo[5.4.0]undec-7-ene), DCC(N,N′-dicyclohexylcarbodiimide), DCE (1,2-dichloroethane), DCM(dichloromethane), DEAD (diethylazodicarboxylate), DIEA(diisopropylethylamine), DMA (N,N-dimethylacetamide), DMAP(4-(N,N-dimethylamino)pyridine), DMF (N,N-dimethylformamide), DMSO(dimethyl sulfoxide), EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride), Et (ethyl), EtOAc (ethyl acetate), EtOH (ethanol), HATU(O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate), HBTU(O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate),HF (hydrogen fluoride), HOAc (acetic acid), HOBT (1-hydroxybenzotriazolehydrate), HPLC (high pressure liquid chromatography), IPA (isopropylalcohol), KHMDS (potassium bis(trimethylsilyl)amide), KN(TMS)₂(potassium bis(trimethylsilyl)amide), KO^(t)Bu (potassiumtert-butoxide), LDA (lithium diisopropylamine), MCPBA(3-chloroperbenzoic acid), Me (methyl), MeCN (acetonitrile), MeOH(methanol), NaBH(OAc)₃ (sodium triacetoxyborohydride), NaCNBH₃ (sodiumcyanoborohydride), NaH (sodium hydride), NaN(TMS)₂ (sodiumbis(trimethylsilyl)amide), NaOAc (sodium acetate), NaOEt (sodiumethoxide), Phe (phenylalanine), PPTS (pyridinium p-toluenesulfonate), PS(polymer supported), Py (pyridine), pyBOP(benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate),TEA (triethylamine), TFA (trifluoroacetic acid), TFAA (trifluoroaceticanhydride), THF (tetrahydrofuran), TLC (thin layer chromatography), Tol(toluoyl), Val (valine), and the like.

Scheme 1 provides a general procedure that can be used to preparesaturated 5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

The saturated cyclic N-substituted-β-amino acid ester intermediates,which can be obtained as described by one of the methods in schemes 3,4, 6, 7 or 8 can be condensed with a carboxylic acid intermediate (or asalt thereof, e.g., sodium salt) using standard peptide couplingconditions used for the formation of amide bonds, such as DCC, to yieldthe shown amide. This intermediate can be cyclized with or withoutisolation in the presence of a base (e.g., triethylamine) to give thedesired saturated 5,6-dihydro-1H-pyridin-2-one compounds.

Scheme 2 provides a general procedure that can be used to prepareunsaturated 5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

The unsaturated cyclic N-substituted-β-amino acid ester intermediates(with R as defined in scheme 1), which can be obtained as described byone of the methods in schemes 5 or 9, can be condensed with a carboxylicacid intermediate (or a salt thereof, e.g., sodium salt) using standardpeptide coupling conditions used for the formation of amide bonds, suchas DCC, to yield the shown amide. This intermediate can be cyclized withor without isolation in the presence of a base (e.g., triethylamine) togive the desired unsaturated 5,6-dihydro-1H-pyridin-2-one compounds.

Scheme 3 provides a general procedure that can be used to preparesaturated cyclic N-substituted-β-amino acid ester intermediates fromsaturated anhydrides.

Commercially available saturated cyclic meso-anhydrides can bedesymmetrized with the help of enzymes or chiral reagents, such ascinchona alkaloids (e.g., quinine or quinidine) as described in theliterature to provide optically active saturated cyclic dicarboxylicacid monoesters (with R as defined in scheme 1). See J. Org. Chem., 65,6984-6991 (2000); Synthesis, 11, 1719-1730 (2001), and references citedtherein.

These intermediates can be further elaborated into protected opticallyactive saturated cyclic β-amino acid esters (e.g., Cbz-protected) via arearrangement reaction, such as the Curtius rearrangement (shown) or aHofmann degradation. Hydrogenation of the protected saturated cyclicβ-amino acid esters under standard conditions can be used to remove theprotecting group and furnish the optically active saturated cyclicβ-amino acid esters, which can be isolated (and used) as either the freebases or their corresponding salts. The optically active saturatedcyclic β-amino acid esters (or their salts) can then be treated withaldehydes or ketones, where R^(x) and R^(w) are independently C₁-C₅alkyl, C₃-C₈ cycloalkyl, —C₁-C₅ alkylene(C₃-C₈ cycloalkyl), —C₁-C₅alkylene(aryl), —C₁-C₅ alkylene(heterocyclyl), aryl, or heterocyclyl, orR^(w) can combine with R^(x) to form a 3- to 8-membered ring, in thepresence of a reducing agent (such as sodium cyanoborohydride) to affordthe desired optically active saturated cyclic N-substituted-β-amino acidester intermediates. Alternatively, the reaction sequence describedabove can be performed without enzymes or chiral reagents leading to thecorresponding achiral intermediates and products.

Scheme 4 provides a general procedure that can be used to preparesaturated cyclic N-substituted-β-amino acid ester intermediates fromunsaturated anhydrides.

Commercially available unsaturated cyclic meso-anhydrides can bedesymmetrized as described above (scheme 3) to provide optically activeunsaturated cyclic dicarboxylic acid monoesters (with R as defined inscheme 1). These intermediates can be further elaborated into protectedoptically active unsaturated cyclic β-amino acid esters (e.g.,Cbz-protected) via a rearrangement reaction, such as the Curtiusrearrangement (shown) or a Hofmann degradation. Hydrogenation of theprotected optically active unsaturated cyclic β-amino acid esters understandard conditions removes the protecting group and reduces the olefinto furnish the optically active saturated cyclic β-amino acid esters,which can be isolated (and used) as either the free bases or theircorresponding salts.

The optically active saturated cyclic β-amino acid esters (or theirsalts) can then be treated with aldehydes or ketones (with R^(x) andR^(w) as defined in scheme 3) in the presence of a reducing agent (suchas sodium cyanoborohydride) to afford the desired optically activesaturated cyclic N-substituted-β-amino acid ester intermediates.Alternatively, the reaction sequence described above can be performedwithout enzymes or chiral reagents leading to the corresponding achiralintermediates and products.

Scheme 5 provides a general procedure that can be used to prepareunsaturated cyclic N-substituted-β-amino acid ester intermediates fromunsaturated anhydrides.

Commercially available unsaturated cyclic meso-anhydrides can bedesymmetrized as described above (scheme 4) to provide optically activeunsaturated cyclic dicarboxylic acid monoesters (with R as defined inscheme 1). These intermediates can be further elaborated into protectedoptically active unsaturated cyclic β-amino acid esters (e.g.,Boc-protected) via a rearrangement reaction, such as the Curtiusrearrangement (shown) or a Hofmann degradation. The Boc protecting groupcan then be selectively removed in the presence of the olefin, thusleading to the optically active unsaturated cyclic β-amino acid esterintermediates, which can be isolated (and used) as either the salts ortheir corresponding free bases.

The optically active unsaturated cyclic β-amino acid esters (or theirsalts) can then be treated with aldehydes or ketones (with R^(x) andR^(w) as defined in scheme 3) in the presence of a reducing agent (suchas sodium cyanoborohydride) to afford the desired optically activeunsaturated cyclic N-substituted-β-amino acid ester intermediates.Alternatively, the reaction sequence described above can be performedwithout enzymes or chiral reagents leading to the corresponding achiralintermediates and products.

Scheme 6 provides an alternate general procedure that can be used toprepare saturated cyclic N-substituted-β-amino acid ester intermediates.

Bicyclic olefins, such as norbornene, can be reacted with chlorosulfonylisocyanate to yield the β-lactams shown. These intermediates can behydrolyzed in the presence of a strong acid (such as hydrochloric acid)to afford the saturated cyclic β-amino acids (or their salts), which canthen be further elaborated into the corresponding esters using standardconditions (with R as defined in scheme 1). The saturated cyclic β-aminoacid esters can then be treated with aldehydes or ketones (with R^(x)and R^(w) as defined in scheme 3) in the presence of a reducing agent,such as sodium cyanoborohydride, to afford the desired saturated cyclicN-substituted-β-amino acid ester intermediates.

Scheme 7 provides a general scheme describing a method that can be usedto resolve the di-exo enantiomers by diastereomeric crystallization

The racemic di-exo-β-amino acid ester derivatives obtained fromnorbornene as described above, can be resolved by forming diastereomericsalts with an optically pure acid, such as (1S)-(+)-10-camphorsulfonicacid. The (1R,2R,3S,4S)-β-amino acid ester derivatives form acrystalline salt with (1S)-(+)-10-camphorsulfonic acid that can beselectively isolated by filtration from an appropriate solvent (e.g.,ethyl acetate) and treated with a base, such as sodium carbonate, toafford the free enantiomerically pure cyclic (1R,2R,3S,4S)-β-amino acidesters. The optically pure cyclic (1R,2R,3S,4S)-3-amino acid esters (ortheir salts) can then be treated with aldehydes or ketones (with R^(x)and R^(w) as defined in scheme 3) in the presence of a reducing agent,such as sodium cyanoborohydride, to afford the desired optically puresaturated cyclic N-substituted-(1R,2R,3S,4S)-β-amino acid esterintermediates.

Scheme 8 provides an alternative procedure that can be used to prepareenantiomerically pure saturated cyclic N-substituted-β-amino acid esterintermediates.

The β-lactam (prepared as described in scheme 6) can be opened andresolved by forming diastereomeric salts with an optically pure acid,such as (1S)-(+)-10-camphorsulfonic acid (as described in scheme 7) inthe presence of an alcohol (e.g., ethanol) to directly afford thediastereomerically pure (1R,2R,3S,4S)-β-amino acid ester as a salt with(1S)-(+)-10-camphorsulfonic acid. Treatment with a base, such aspotassium carbonate, followed by reductive alkylation with aldehydes orketones (with R^(x) and R^(w) as defined in scheme 3) in the presence ofa reducing agent, such as sodium cyanoborohydride, affords the desiredenantiomerically pure saturated cyclicN-substituted-(1R,2R,3S,4S)-β-amino acid ester intermediates.

Scheme 9 provides a general procedure that can be used to preparesaturated cyclic N-substituted-β-amino acid ester intermediates.

The saturated cyclic β-amino acid esters (or their salts, with R asdefined in scheme 1) can be purchased, prepared from the correspondingcommercially available saturated cyclic β-amino acids, or can beprepared by methods described in schemes 3, 4, 6 or 7. The saturatedcyclic β-amino acid esters can then be treated with aldehydes or ketones(with R^(x) and R^(w) as defined in scheme 3) in the presence of areducing agent, such as sodium cyanoborohydride, to afford the desiredsaturated cyclic N-substituted-β-amino acid ester intermediates. In eachcase the saturated cyclic β-amino acid esters or the desired saturatedcyclic N-substituted-β-amino acid ester intermediates may be opticallyactive.

Scheme 10 provides an alternative general procedure that can be used toprepare saturated cyclic N-substituted-β-amino acid ester intermediates.

The saturated cyclic β-amino acid esters (or their salts, with R asdefined in scheme 1) can be purchased, prepared from the correspondingcommercially available saturated cyclic β-amino acids, or can beprepared by the method described in schemes 3, 4, 6 or 7. The saturatedcyclic β-amino acid esters can then be treated with halides orpseudohalides X^(a) (e.g., bromides, iodides or triflates), where R^(v)is aryl or heterocyclyl, in the presence of metal catalyst such ascopper (e.g., under Ullmann reaction conditions) or palladium (e.g.,under Buchwald-Hartwig reaction conditions), to afford the desiredsaturated cyclic N-substituted-β-amino acid ester intermediates. In eachcase the saturated cyclic β-amino acid esters or the desired saturatedcyclic N-substituted-β-amino acid ester intermediates may be opticallyactive.

Scheme 11 provides a general procedure that can be used to prepareunsaturated cyclic N-substituted-β-amino acid ester intermediates.

The unsaturated cyclic β-amino acid esters (or their salts, with R asdefined in scheme 1) can be purchased, prepared from the correspondingcommercially available unsaturated cyclic β-amino acids, or can beprepared by the method described in scheme 5. The unsaturated cyclicβ-amino acid esters can then be treated with aldehydes or ketones (withR^(x) and R^(w) as defined in scheme 3) in the presence of a reducingagent, such as sodium cyanoborohydride, to afford the desiredunsaturated cyclic N-substituted-β-amino acid ester intermediates. Ineach case the unsaturated cyclic β-amino acid esters or the desiredunsaturated cyclic N-substituted-β-amino acid ester intermediates may beoptically active.

Scheme 12 provides an alternative general procedure that can be used toprepare unsaturated cyclic N-substituted-β-amino acid esterintermediates.

The unsaturated cyclic β-amino acid esters (or their salts, with R asdefined in scheme 1) can be purchased, prepared from the correspondingcommercially available unsaturated cyclic β-amino acids, or can beprepared by the method described in scheme 5. The unsaturated cyclicβ-amino acid esters can then be treated with halides or pseudohalidesX^(a) (e.g., bromides, iodides or triflates), where R^(v) is aryl orheterocyclyl, in the presence of metal catalyst such as copper (e.g.,under Ullmann reaction conditions), to afford the desired unsaturatedcyclic N-substituted-β-amino acid ester intermediates. In each case theunsaturated cyclic β-amino acid esters or the desired unsaturated cyclicN-substituted-β-amino acid ester intermediates may be optically active.

Scheme 13 provides a general procedure that can be used to prepare7-substituted-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl-aceticacid intermediates.

Commercially available 6-nitrobenzothiazole can be treated withhydrazine to obtain the 2-amino-5-nitro-benzenethiol, which cansubsequently be reacted with chloroacetoacetate to give the(7-nitro-4H-benzo[1,4]thiazin-3-yl)-acetic acid ethyl ester. Reductionof the nitro group to the amino group can be accomplished by reactionwith tin(II)chloride. Subsequent reaction with a sulfonyl chloride, suchas methylsulfonyl chloride, can be used to obtain the correspondingsulfonamides. Protection of both nitrogens with a suitable protectinggroup such as a Boc group can be achieved by using standard methods forprotecting amino groups. The sulfides can be oxidized using as suitableoxidizing reagent (e.g. MCPBA) to give the sulfones. Finally,deprotection of the amino groups using trifluoroacetic acid, followed byhydrolysis of the esters can be used to afford the desired acidintermediates.

Scheme 14 provides a general procedure that can be used to prepare7-substituted-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl-aceticacid intermediates.

Commercially available 2-chloro-5-nitro-benzenesulfonic acid can betreated with thionyl chloride to give the sulfonylchloride, which can befurther treated with ammonia to afford the sulfonamide intermediate. Thechloride can be displaced with ammonia by treatment with ammoniumhydroxide and ammonium carbonate in the presence of copper(II)sulfate.Reduction of the nitro group under standard hydrogenation conditionsaffords the aniline intermediate, which can be treated with a sulfonylchloride, such as methylsulfonyl chloride, to yield the correspondingsulfonamide. Acylation of the 2-amino moiety with malonyl chlorides,e.g., ethyl 3-chloro-3-oxo-propionate, gives the corresponding amide,which can simultaneously be cyclized to the thiadiazine-dioxide andhydrolyzed to the desired acid intermediate.

Scheme 15 provides an alternative procedure that can be used to preparethe 2-amino-5-nitro-benzenesulfonamide intermediate.

Commercially available 2-amino-5-nitro-benzenesulfonic acid can beconverted to the corresponding sulfonyl chloride with phosphorylchloride in the presence of a suitable co-solvent, such as sulfolane.Treatment with ammonia, e.g., aqueous ammonium hydroxide solution orammonia gas, affords the desired 2-amino-5-nitro-benzenesulfonamideintermediate.

Scheme 16 provides an alternative procedure that can be used to preparethe 2-amino-5-methanesulfonylamino-benzenesulfonamide intermediate.

The 2-chloro-5-nitro-benzenesulfonamide intermediate (prepared asdescribed in scheme 14) can be treated with a benzylic amine, such asbenzylamine, to displace the chloro moiety. Hydrogenation under standardconditions in the presence of an acid (e.g., methanesulfonic acid) canbe used to remove the benzylic group and to reduce the nitro group atthe same time to afford the 2,5-diamino-benzenesulfonamide intermediateas a salt. Alternatively, the 2,5-diamino-benzenesulfonamide salt can beprepared by reduction of the 2-amino-5-nitro-benzenesulfonamide(prepared as described in schemes 14 and 15) under standardhydrogenation conditions in the presence of an acid (e.g.,methanesulfonic acid). Subsequent reaction with methanesulfonyl chlorideaffords the desired 2-amino-5-methanesulfonylamino-benzenesulfonamideintermediate.

Scheme 17 provides a procedure that was used to prepare the(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid intermediate.

Commercially available 2-aminobenzenesulfonamide can be treated withN-iodosuccinimide (NIS) to afford 2-amino-5-iodo-benzenesulfonamide.Acylation with a malonyl halide monoester, such as ethyl3-chloro-3-oxo-propionate, or with a dialkyl malonate, such as diethylmalonate, affords the corresponding amide, which can simultaneously becyclized to the thiadiazine-dioxide and hydrolyzed to the desired acidintermediate (or a salt thereof, e.g., sodium salt).

Scheme 18 provides a procedure that was used to prepare the(1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetic acidintermediate.

Commercially available 2-aminobenzenesulfonamide can be acylated with amalonyl halide monoester, such as ethyl 3-chloro-3-oxo-propionate, orwith a dialkyl malonate, such as diethyl malonate, to afford thecorresponding amide, which can simultaneously be cyclized to thethiadiazine-dioxide and hydrolyzed to the desired acid intermediate (ora salt thereof, e.g., sodium salt).

Scheme 19 provides a procedure that was used to prepare the3-amino-pyridine-4-sulfonic acid amide intermediate.

The 3-amino-pyridine-4-sulfonic acid amide intermediate can be preparedfollowing procedures described in Tetrahedron 1998, 54, 13645-13654(steps 2-4). Commercially available 4-chloro-3-nitro-pyridine can betreated with sodium sulfide hydrate to displace the chloro moiety.Treatment with chloramine followed by oxidation of the sulfur with anoxidizing agent, such as MCPBA, furnishes the nitro-sulfonamideintermediate. Reduction of the nitro group using standard conditions(e.g., tin(II)chloride) affords the desired 3-amino-pyridine-4-sulfonicacid amide intermediate.

Scheme 20 provides a procedure that was used to prepare the4-amino-pyridine-3-sulfonic acid amide intermediate.

Commercially available pyridin-4-ylamine can be treated withchlorosulfonic acid in the presence of thionyl chloride to afford thesulfonyl chloride intermediate. Treatment with aqueous ammoniumhydroxide solution affords the desired 4-amino-pyridine-3-sulfonic acidamide intermediate.

Scheme 21 provides a procedure that was used to prepare the3-amino-pyridine-2-sulfonic acid amide intermediate.

Commercially available 2-chloro-3-nitro-pyridine can be treated withbenzyl mercaptan in the presence of a base (e.g., potassium carbonate)to afford the sulfide intermediate. Oxidation with1,3-dichloro-5,5-dimethyl-hydantoin furnishes the sulfonyl chloride,which can be treated with aqueous ammonium hydroxide solution to yieldthe sulfonamide intermediate. Reduction of the nitro group understandard hydrogenation conditions affords the desired3-amino-pyridine-2-sulfonic acid amide intermediate.

Scheme 22 provides a procedure that was used to prepare the4-amino-pyridine-3-sulfonic acid amide intermediate.

Commercially available 4-chloro-pyridine-3-sulfonic acid amide can betreated with sodium azide to furnish the azido intermediate, which canbe reduced with a reducing agent (e.g., sodium borohydride) to affordthe desired 4-amino-pyridine-3-sulfonic acid amide intermediate.

Scheme 23 provides a procedure that was used to prepare the acidintermediates derived from aminopyridine sulfonic acid amideintermediates.

The aminopyridine sulfonic acid amide intermediates described in schemes19-21 can be acylated with a malonyl halide monoester, such as ethyl3-chloro-3-oxo-propionate, or with a dialkyl malonate, such as diethylmalonate, to afford the corresponding amide, which can simultaneously becyclized to the thiadiazine-dioxide and hydrolyzed to the desired acidintermediate (or a salt thereof, e.g., sodium salt).

Scheme 24 provides a general procedure that was used to prepare5,6-dihydro-1H-pyridin-2-one compounds of Formula I from thecorresponding iodo precursors.

Optionally substituted 5,6-dihydro-1H-pyridin-2-ones can be treated withsubstituted sulfonamides in a copper-mediated displacement reaction toafford the desired 5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

Scheme 25 provides a general procedure that was used to prepare5,6-dihydro-1H-pyridin-2-one compounds of Formula I bearing a sulfamidemoiety from the corresponding iodo precursors.

Optionally substituted 5,6-dihydro-1H-pyridin-2-ones can be treated withsodium azide to displace the iodo moiety. Reduction of theazido-intermediate using standard conditions, such as catalytichydrogenation, affords the aniline intermediate. Further reaction withan optionally substituted sulfamoyl chloride in the presence of a base(e.g., triethylamine) affords the desired 5,6-dihydro-1H-pyridin-2-onecompounds of Formula I.

Scheme 26 provides a general procedure that was used to prepareN-substituted 5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

The sulfonamide moiety of optionally substituted5,6-dihydro-1H-pyridin-2-ones can be N-alkylated by treatment with analkylating agent, such as alkyl halides or pseudohalides X^(a) (e.g.,chlorides, bromides, iodides, mesylates, tosylates, triflates, orchloroformates), in the presence of a base (e.g., potassium carbonate)to afford the desired N-substituted 5,6-dihydro-1H-pyridin-2-onecompounds of Formula I.

Scheme 27 provides a procedure that was used to prepare the5,6-dihydro-1H-pyridin-2-one compounds of Formula I from thecorresponding iodo precursors.

Optionally substituted 5,6-dihydro-1H-pyridin-2-ones can be treated withstannanes, such as the unsaturated cyclic sulfone shown above, in aStille-type palladium-catalyzed reaction to afford the unsaturatedintermediates shown. Reduction of the alkene using standardhydrogenation conditions affords the desired5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

Scheme 28 provides a general procedure that was used to prepare the5,6-dihydro-1H-pyridin-2-one compounds of Formula I from thecorresponding iodo precursors.

Optionally substituted 5,6-dihydro-1H-pyridin-2-ones can be treated withamines in a copper-mediated displacement reaction to afford the desired5,6-dihydro-1H-pyridin-2-one compounds of Formula I.

Example 1(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) 2-Chloro-5-nitrobenzenesulfonamide

To a solution of thionyl chloride (11 mL) and2-chloro-5-nitro-benzenesulfonic acid (4.78 g, 20.1 mmol) was addedN,N-dimethylformamide (0.92 μL) and the reaction mixture was heated atreflux for 4 h. The reaction mixture was then carefully quenched bypouring it into water and the product was isolated by vacuum filtration.The sulfonyl chloride was dissolved in a minimal amount of toluene andthen added to a mixture of concentrated aqueous ammonium hydroxidesolution (25 mL) and tetrahydrofuran (25 mL) at −10° C. After stirringfor 2 h the reaction was quenched by adding a 6.0M aqueous hydrochloricacid solution until pH 4 was reached. The layers were separated and theorganic layer was concentrated in vacuo to a slurry. Pentane was addedand the product was isolated by vacuum filtration to afford2-chloro-5-nitrobenzenesulfonamide (2.0 g, 8.48 mmol, 42.4%), as asolid. ¹H NMR (400 MHz, DMSO-d₆) δ: 7.94 (d, 1H, J=8.8 Hz), 7.97 (bs,2H), 8.40 (dd, 1H, J₁=8.6 Hz, J₂=3.1 Hz), 8.64 (d, 1H, J=3.1 Hz).

b) 2-Amino-5-nitrobenzenesulfonamide

2-Chloro-5-nitro-benzenesulfonamide (1.95 kg, 8.30 mol), ammoniumcarbonate (1.983 kg, 20.64 mol), and copper (II) sulfate (394 g, 2.47mol) were charged to an autoclave and diluted with a 30% aqueousammonium hydroxide solution (11.7 L, 330 mol). The mixture was heated at118° C. for 3 days and was then cooled to 23° C. The mixture wasfiltered and the solids were then washed with water (20 L). This solidwas dissolved in hot methanol (20 mL/g), and the mixture was filtered toremove undissolved solids. The filtrate was stored at 4° C. overnight,and the resulting solid product was then filtered. The filtrate waspartially concentrated by vacuum distillation and, when the concentratewas cooled to 23° C., the solid product was then filtered off. The twocrops of solid were combined and further dried in vacuo at 45° C. toafford the desired product, 2-amino-5-nitro-benzenesulfonamide (1.10 kg,5.06 mol, 61%), as a solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 6.89 (d, J=9.3Hz, 1H), 7.12 (bs, 2H), 7.57 (bs, 2H), 8.07 (dd, J₁=9.0 Hz, J₂=2.6 Hz,1H), 8.43 (d, J=3.0 Hz, 1H).

Alternatively, 2-amino-5-nitrobenzenesulfonamide can be prepared asfollows:

2-Amino-5-nitrobenzenesulfonic acid (200.00 g, 0.917 mol) was suspendedin warm sulfolane (250 mL) and the suspension was heated to 80° C.Phosphorous oxychloride (126 mL, 1.375 mol) was added and resultingmixture was heated to 110-120° C. and stirred for 4 h. The resultingsolution was cooled to 60° C. and added dropwise into concentratedaqueous ammonium hydroxide solution (800 mL, 11.9 mol) at <10° C. Theflask was rinsed with warm sulfolane (50 mL) and the wash was added intothe above reaction mixture. The resulting suspension was stirred at 25°C. for 1 h, heated to 95° C. and stirred for 1 hour. The mixture wascooled to 80° C. and the pH was adjusted to 6-8 with 3.0M aqueoushydrochloric acid solution (˜600 mL) and allowed to cool to 25° C. Thedark green suspension was filtered, and the wet filter cake was washedwith water (300 mL) and dried at 60° C. overnight to give the crudeproduct (140 g) as a green-yellow solid. The crude product was dissolvedin 0.5M aqueous sodium hydroxide solution (1.4 L, 0.7 mol). Charcoal (14g) was added and the mixture was heated to reflux and stirred for 15min. The mixture was filtered through Celite and washed with 0.5Maqueous sodium hydroxide solution (100 mL). The pH of the filtrate wasadjusted to 6-8 with concentrated aqueous hydrochloric acid solution(˜60 mL) and the yellow suspension was allowed to cool to 25° C. Themixture was filtered and the wet filter cake was washed with water (200mL) and dried at 60° C. overnight to afford the desired product,2-amino-5-nitrobenzenesulfonamide (130 g, 0.599 mol, 65%) as a brightyellow powder.

c) 2,5-Diaminobenzenesulfonamide

2-Amino-5-nitro-benzenesulfonamide (5.00 kg, 23.0 mol), methanol (65 L),tetrahydrofuran (65 L), and 10% palladium on carbon (250 g) were chargedto an autoclave. The mixture was cycled with nitrogen and hydrogenpurges (3×), and the mixture was then stirred under hydrogen (50 psi) at23° C. overnight. The catalyst was removed by filtration and thefiltrate was then concentrated in vacuo to give a brown solid. The solidwas further dried in vacuo at 45° C. to afford the desired product,2,5-diamino-benzenesulfonamide (4.21 kg, 22.4 mol, 98%), as a solid. ¹HNMR (400 MHz, DMSO-d₆) δ: 4.54 (2H, bs), 4.98 (2H, bs), 6.55-6.60 (2H,m), 6.87 (1H, d, J=2.2 Hz), 6.99 (2H, bs). LC-MS (ESI) calcd forC₆H₉N₃O₂S 187.04, found 188.3 [M+H⁺].

d) 2-Amino-5-methanesulfonylamino-benzenesulfonamide

2,5-Diamino-benzenesulfonamide (4.20 kg, 22.4 mol) was dissolved indichloromethane (120 L) and pyridine (8.00 kg, 89.9 mol), and theresulting solution was cooled to 0° C. Methanesulfonyl chloride (2.80kg, 24.4 mol) was added slowly, and the resulting mixture was allowed towarm to 23° C. and stirred for 2 days. The mixture was filtered and theresulting solid was washed with dichloromethane (2×20 L). The solid wasdiluted with water (100 L) and 1.0M aqueous hydrochloric acid solution(25 L), and was then stirred at 23° C. for 1 h. The mixture was filteredand the resulting solid was washed with water (20 L) and then withmethyl-tert-butyl ether (2×10 L). The solid was further dried in vacuoat 45° C. to afford the desired product,2-amino-5-methanesulfonylamino-benzenesulfonamide (4.39 kg, 16.5 mol,73%) as a pale pink solid. ¹H NMR (400 MHz, CD₃OD) δ: 2.89 (3H, s), 6.82(1H, d, J=8.5 Hz), 7.20 (1H, dd, J₁=8.5 Hz, J₂=2.5 Hz), 7.58 (1H, d,J=2.5 Hz). LC-MS (ESI) calcd for C₇H₁₁N₃O₄S₂ 265.02, found 266.0 [M+H⁺].

Alternatively, 2-amino-5-methanesulfonylamino-benzenesulfonamide can beprepared as follows:

a′) 2-Benzylamino-5-nitro-benzenesulfonamide

A mixture of 2-chloro-5-nitro-benzenesulfonamide (2.20 kg, 9.30 mol),benzylamine (1.5 L, 13.9 mol), triethylamine (2.5 L, 18.1 mol), andacetonitrile (22.0 L) were heated at 92° C. for 20 h. The mixture wasthen cooled to 40° C., and was then partially concentrated in vacuo. Theresidue was added to 0° C. water (22.0 L) and the resulting suspensionwas allowed to warm to 23° C. and stirred for 2 h. The suspension wasfiltered and the solid was then washed with water (5 L). The washedsolid was suspended in absolute ethanol (11 L), and was then filteredand washed with absolute ethanol (5 L). The solid was further dried invacuo at 45° C. to afford the desired product,2-benzylamino-5-nitro-benzenesulfonamide (2.40 kg, 7.81 mol, 84%), as ayellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 4.64 (2H, d, J=4.6 Hz), 6.81(1H, d, J=9.4 Hz), 7.23-7.44 (6H, m), 7.77 (2H, bs), 8.11 (1H, dd,J₁=9.4 Hz, J₂=2.3 Hz), 8.49 (1H, d, J=3.1 Hz). LC-MS (ESI) calcd forC₁₃H₁₃N₃O₄S 307.06, found 308.2 [M+H⁺] (100%), 615.2 [2M+H⁺] (81%).

b′) 2,5-Diamino-benzenesulfonamide methanesulfonate

Methanesulfonic acid (465 mL, 7.16 mol) was added slowly to a solutionof 2-benzylamino-5-nitro-benzenesulfonamide (2.20 kg, 7.16 mol) andtetrahydrofuran (11.0 L). The resulting solution was added to a mixtureof 10% palladium on carbon (220 g of 50% water wet catalyst) and water(1.1 L) in a hydrogenation reactor. The mixture was further diluted withabsolute ethanol (21.0 L) and was then hydrogenated with 55 psi hydrogenat 50° C. for 21 h. Additional 10% palladium on carbon (55 g of 50%water wet catalyst) was added, and hydrogenation at 55 psi and 50° C.was continued for 22 h. The resulting suspension was diluted with water(1.1 L) and the suspension was then filtered through a pad of Celite.The filtrate was partially concentrated in vacuo and was then dilutedwith acetonitrile (15.4 L). The solution was again partiallyconcentrated in vacuo and diluted with acetonitrile (15.4 L). Theresulting suspension was partially concentrated in vacuo and was allowedto stir at 23° C. for 2 h. The suspension was filtered and the solid wasthen washed with acetonitrile (3 L). The solid was further dried invacuo at 45° C. to afford the desired product,2,5-diamino-benzenesulfonamide methanesulfonate (1.88 kg, 6.64 mol,93%), as a purple solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 2.34 (3H, s), 6.05(2H, b), 6.87 (1H, d, J=8.6 Hz), 7.20 (1H, dd, J₁=8.6 Hz, J₂=2.3 Hz),7.38 (2H, s), 7.53 (1H, d, J=2.3 Hz), 9.62 (3H, b). LC-MS (ESI) calcdfor C₆H₉NO₂S 187.04, found 187.9 [M+H⁺].

Alternatively, 2,5-diamino-benzenesulfonamide methanesulfonate can beprepared as follows:

2-Amino-5-nitrobenzenesulfonamide (prepared as described in Example 1b,100.00 g, 0.460 mol) and 5% palladium on carbon (wet, 5.00 g) weresuspended in ethanol (2 L) and water (100 mL). Methanesulfonic acid (33mL, 0.51 mol) was added, and the resulting mixture was heated to 55° C.and stirred under atmospheric hydrogen for 8 h. The mixture was filteredand the filtrate was concentrated in vacuo to a volume of about 450 mL.To the concentrate was added acetonitrile (1 L) and resulting mixturewas stirred at 25° C. overnight. The suspension was filtered to affordthe desired product, 2,5-diamino-benzenesulfonamide methanesulfonate(122.36 g, 0.432 mol, 93.8%) as a purple solid.

c′) 2-Amino-5-methanesulfonylamino-benzenesulfonamide

2,5-Diamino-benzenesulfonamide methanesulfonate (1.80 kg, 6.35 mol) wassuspended in acetonitrile (24 L). Pyridine (1.55 L, 19.1 mol) was added,followed by the careful slow addition of methanesulfonyl chloride (517mL, 6.68 mol). After stirring at 23° C. for 20 h, the mixture waspartially concentrated in vacuo at 55° C. Water (18 L) was added to theconcentrate, and the resulting suspension was stirred at 23° C. for 2 h.The solid was filtered and was then washed with water (4 L) and airdried on the filter. The solid was suspended in absolute ethanol (9 L),stirred at 23° C. for 9 h, and was then filtered. The solid was washedwith absolute ethanol (2×2 L), and was then further dried in vacuo at50° C. to afford the desired product,2-amino-5-methanesulfonylamino-benzenesulfonamide (1.45 kg, 5.48 mol,86%), as a purple solid.

e) N-(4-Methanesulfonylamino-2-sulfamoyl-phenyl)-malonamic acid ethylester

2-Amino-5-methanesulfonylamino-benzenesulfonamide (23.27 g, 87.81 mmol)was dissolved in N,N-dimethylacetamide (100 mL) and diethyl ether (100mL). Ethyl 3-chloro-3-oxo-propionate (13.88 g, 92.20 mmol) was added andthe reaction mixture was stirred at 25° C. for 1 h. The reaction mixturewas diluted with ethyl acetate (400 mL) and was extracted with water(400 mL). The aqueous layer was back-extracted with ethyl acetate (2×200mL). The combined organic layers were dried over sodium sulfate,filtered, and most of the solvent was removed in vacuo to a volume of˜100 mL.

To the stirred solution was added hexanes (˜100 mL) upon which aprecipitate formed. The precipitate was collected by vacuum filtration,washed with hexanes and dried under high vacuum to afford theanalytically pure product,N-(4-methanesulfonylamino-2-sulfamoyl-phenyl)-malonamic acid ethyl ester(31.22 g, 85.53 mmol, 97.4%), as a light-brown solid. ¹H NMR (400 MHz,CD₃OD) δ: 1.31 (3H, t, J=7.0 Hz), 3.00 (3H, s), 3.59 (2H, s), 4.25 (2H,quartet, J=6.9 Hz), 7.42-7.45 (1H, m), 7.86 (1H, m), 7.92 (1H, d, J=8.8Hz).

f) N-(4-Methanesulfonylamino-2-sulfamoylphenyl)-malonamic acid methylester

2-Amino-5-methanesulfonylamino-benzenesulfonamide (prepared as describedin Example 1d, 1.70 kg, 6.40 mol) was dissolved in tetrahydrofuran (35L), and was then cooled to 0° C. Methyl 3-chloro-3-oxopropionate (792mL, 7.40 mol) was added slowly, and the resulting mixture was thenallowed to warm to 23° C. and stirred for 2 days. The solvent wasremoved in vacuo, and the residue was then diluted with water (4 L) andsaturated aqueous sodium bicarbonate solution (2 L). The resulting solidwas filtered, and was then washed with water (5 L). The solid wassuspended in hot methanol (15 mL/g), and was then cooled to 23° C. andfiltered to afford the desired product,N-(4-methanesulfonylamino-2-sulfamoyl-phenyl)-malonamic acid methylester (1.68 kg, 4.61 mol, 72%), as a brown solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 3.02 (3H, s), 3.60 (2H, s), 3.66 (3H, s), 7.38 (1H, dd,J₁=2.3 Hz, J₂=8.6 Hz), 7.53 (2H, bs), 7.73 (1H, d, J=2.4 Hz), 7.83 (1H,d, J=8.7 Hz), 9.43 (1H, s), 9.99 (1H, s).

g)(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid

N-(4-Methanesulfonylamino-2-sulfamoyl-phenyl)-malonamic acid ethyl ester(prepared as described in Example 1e, 9.55 g, 26.16 mmol) was dissolvedin 8% aqueous sodium hydroxide solution (262 mL) and heated at 100° C.for 1.5 h. The reaction mixture was cooled to 0° C. and the solution wasacidified by slowly adding 12.0M aqueous hydrochloric acid solutionuntil pH 1-2 was reached. A precipitate started to form and thesuspension was allowed to stir for 30 min at 0° C. The precipitate wascollected by vacuum filtration, washed with cold water, and dried underhigh vacuum to afford the desired product,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetic acid (7.20 g, 21.621mmol, 82.6%), as a pinkish solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 3.03 (3H,s), 3.56 (2H, s), 7.33 (1H, d, J=9.1 Hz), 7.52-7.54 (2H, m), 10.09 (1H,s), 12.24 (1H, s), 13.02 (1H, bs). LC-MS (ESI) calcd for C₁₀H₁₁N₃O₆S₂333.01, found 334.1 [M+H⁺].

Alternatively,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid can be prepared as follows:

N-(4-methanesulfonylamino-2-sulfamoyl-phenyl)-malonamic acid methylester (prepared as described in Example 1g, 1.35 kg, 3.69 mol) was addedto 3.8 wt. % aqueous sodium hydroxide solution (14.0 kg). The resultingmixture was stirred at 23° C. for 30 h, and was then cooled to 0° C. A2.0M aqueous hydrochloric acid solution (9.72 L) was slowly added,stirring at 0° C. was continued for 30 min, and the mixture was thenfiltered. The solid was washed with water (1.4 L), and was then slurriedin a mixture of methanol (1.4 L) and diethyl ether (2.7 L). Afterfiltration, the solid was washed with diethyl ether (2×1.4 L) and wasfurther dried in vacuo at 23° C. to afford the desired product,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (1.07 kg, 3.21 mol, 87%), as a light brown solid.

h)(rac-di-exo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(rac-di-exo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethyl esterhydrochloride (1 g, 4.6 mmol) was suspended in methanol (23 mL). Sodiumacetate (0.755 g, 9.2 mmol) was added followed by 4 Å powdered molecularsieves (1g) and 4-fluoro-benzaldehyde (0.571 g, 4.6 mmol). Sodiumcyanoborohydride (0.578 g, 9.2 mmol) was added and the mixture wasstirred at 25° C. for 16 h. The mixture was poured into a 1:1 mixture ofsaturated aqueous sodium bicarbonate solution (200 mL) and ethyl acetate(200 mL). After shaking, both layers were passed through a plug ofCelite. The organic layer was further washed with saturated aqueousbrine solution (50 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(rac-di-exo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.993 g, 3.41 mmol, 74%), as a clear oil. LC-MS (ESI)calcd for C₁₇H₂₂FNO₂ 291.16, found 292.1 [M+H⁺].

i)(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.299 mmol) wasdissolved in anhydrous N,N-dimethylformamide (1.5 mL).(rac-di-exo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.087 g, 0.3 mmol) was added followed by a 1.0Msolution of N,N-dicyclohexylcarbodiimide in dichloromethane (0.315 mL,0.315 mmol). The mixture was stirred at 25° C. for 1.5 h. Triethylamine(0.124 mL, 0.9 mmol) was added and the mixture was stirred at 50° C. for16 h. Upon cooling, the mixture was poured into 1.0M aqueoushydrochloric acid solution (100 mL). The product was extracted intoethyl acetate (100 mL). The organic layer was washed with 1.0M aqueoushydrochloric acid solution (50 mL), saturated aqueous brine solution (25mL) and dried over magnesium sulfate. Purification by flash columnchromatography (Merck silica gel 60, 40-63 μm, 0 to 1% methanol indichloromethane) followed by crystallization from methanol afforded thedesired product,(rac-di-exo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1)⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.08 g, 0.141 mmol, 47%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.18-1.21 (2H, m), 1.39-1.59 (5H, m), 2.61-2.64 (1H, m), 3.03 (1H, d,J=14.2 Hz), 3.05 (3H, s), 3.53 (1H, d, J=9.3 Hz), 4.41 (1H, d, J=14.8Hz), 4.96 (1H, d, J=15.5 Hz), 7.14 (2H, t, J=9.0 Hz), 7.32 (2H, dd,J₁=8.7 Hz, J₂=6.2 Hz), 7.50 (1H, dd, J₁=9.3 Hz, J₂=2.4 Hz), 7.55-7.57(2H, m), 10.17 (1H, s). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₆S₂ 560.12,found 561.3 [M+H⁺]. HPLC-analysis: Chiralpak AS-RH 4.6×250 mm, 5 micronat r.t., Solvent A-Solvent B (see table for gradient), 0.8 mL/min, 310nm, t1=7.72 min, t2=9.00 min.

Example 2(rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-endo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(rac-di-endo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester hydrochloride (1 g, 4.6 mmol) was suspended in methanol (23 mL).Sodium acetate (0.755 g, 9.2 mmol) was added followed by 4 Å powderedmolecular sieves (1g) and 4-fluoro-benzaldehyde (0.571 g, 4.6 mmol).Sodium cyanoborohydride (0.578 g, 9.2 mmol) was added and the mixturewas stirred at 25° C. for 16 h. The mixture was poured into a 1:1mixture of saturated aqueous sodium bicarbonate solution (200 mL) andethyl acetate (200 mL). After shaking, both layers were passed through aplug of Celite. The organic layer was further washed with saturatedaqueous brine solution (50 mL), dried over magnesium sulfate, filtered,and concentrated in vacuo to afford the crude product,(di-endo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (1.096 g, 3.77 mmol, 82%), as a clear oil. LC-MS (ESI)calcd for C₁₇H₂₂FNO₂ 291.16, found 292.1 [M+H⁺].

b)(rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.299 mmol) wasdissolved in anhydrous N,N-dimethylformamide (1.5 mL).(rac-di-endo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.087 g, 0.3 mmol) was added followed by a 1.0Msolution of N,N-dicyclohexylcarbodiimide in dichloromethane (0.315 mL,0.315 mmol). The mixture was stirred at 25° C. for 1.5 h. Triethylamine(0.124 mL, 0.9 mmol) was added and the mixture was stirred at 50° C. for16 h. Upon cooling, the mixture was poured into 1.0M aqueoushydrochloric acid solution (100 mL). The product was extracted intoethyl acetate (100 mL). The organic layer was washed with 1.0M aqueoushydrochloric acid solution (50 mL), saturated aqueous brine solution (25mL) and dried over magnesium sulfate. Purification by flash columnchromatography (Merck silica gel 60, 40-63 μm, 0 to 1% methanol indichloromethane) followed by crystallization from methanol afforded thedesired product,(rac-di-endo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.092 g, 0.164 mmol, 55%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.25-1.48 (6H, m), 2.64-2.73 (2H, m), 3.06 (3H, s), 3.24 (1H, d,J=23.8 Hz), 3.72 (1H, d, J=11.6 Hz), 4.07 (1H, d, J=14.8 Hz), 5.12 (1H,d, J=15.3 Hz), 7.14 (2H, t, J=8.6 Hz), 7.39 (2H, dd, J₁=8.1 Hz, J₂=5.7Hz), 7.51 (1H, dd, J₁=8.5 Hz, J₂=2.4 Hz), 7.57-7.60 (2H, m), 10.18 (1H,s). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₆S₂ 560.12, found 561.4 [M+H⁺].HPLC-analysis: Chiralpak AS-RH 4.6×250 mm, 5 micron at r.t., SolventA-Solvent B (see table for gradient), 0.8 mL/min, 310 nm, t1=7.58 min,t2=10.08 min.

Example 3(rac-di-endo)-N-{3-[3-(5-Fluoro-pyridin-2-ylmethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-endo)-3-[(5-Fluoro-pyridin-2-ylmethyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(rac-di-endo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester hydrochloride (1 g, 4.6 mmol) was suspended in methanol (23 mL).Sodium acetate (0.755 g, 9.2 mmol) was added followed by 4 Å powderedmolecular sieves (1g) followed by 5-fluoro-pyridine-2-carbaldehyde(0.576 g, 4.6 mmol). Sodium cyanoborohydride (0.578 g, 9.2 mmol) wasadded and the mixture was stirred at 25° C. for 16 h. The mixture waspoured into a 1:1 mixture of saturated aqueous sodium bicarbonatesolution (200 mL) and ethyl acetate (200 mL). After shaking, both layerswere passed through a plug of Celite. The organic layer was furtherwashed with saturated aqueous brine solution (50 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford thecrude product,(rac-di-endo)-3-[(5-fluoro-pyridin-2-ylmethyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (1.247 g, 4.27 mmol, 93%), as a clear oil. LC-MS (ESI)calcd for C₁₆H₂₁FN₂O₂ 292.16, found 293.1 [M+H⁺].

b)(rac-di-endo)-N-{3-[3-(5-Fluoro-pyridin-2-ylmethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.299 mmol) wasdissolved in anhydrous N,N-dimethylformamide (1.5 mL).(rac-di-endo)-3-[(5-Fluoro-pyridin-2-ylmethyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.088 g, 0.3 mmol) was added followed by a 1.0Msolution of N,N-dicyclohexylcarbodiimide in dichloromethane (0.315 mL,0.315 mmol). The mixture was stirred at 25° C. for 1.5 h. Triethylamine(0.124 mL, 0.9 mmol) was added and the mixture was stirred at 50° C. for16 h. Upon cooling, the mixture was poured into 1.0M aqueoushydrochloric acid solution (100 mL). The product was extracted intoethyl acetate (100 mL). The organic layer was washed with 1.0M aqueoushydrochloric acid solution (50 mL), saturated aqueous brine solution (25mL), dried over magnesium sulfate, and concentrated in vacuo.Crystallization from methanol afforded the desired product,(rac-di-endo)-N-{3-[3-(5-fluoro-pyridin-2-ylmethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.087 g, 0.156 mmol, 52%), as a light yellow powder. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.24-1.50 (6H, m), 2.64-2.73 (2H, m), 3.05 (3H, s), 3.28(1H, d, J=12.2 Hz), 3.89 (1H, d, J=14.9 Hz), 4.25 (1H, d, J=15.7 Hz),5.10 (1H, d, J=15.5 Hz), 7.46-7.51 (2H, m), 7.55-7.57 (2H, m), 7.66-7.71(1H, m), 8.49 (1H, d, J=2.3 Hz), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₄H₂₄FN₅O₆S₂ 561.12. found 562.4 [M+H⁺].

Example 4N-{3-[(1S,2S,7R,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1R,2S,3R,4S)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid

The starting material (a) was prepared as described in J. Org. Chem.2000, 65, 6984-6991. cis-5-Norbornene-endo-2,3-dicarboxylic anhydride(4.104 g, 25 mmol) was suspended in a 1:1 mixture of toluene and carbontetrachloride (500 mL). The mixture was stirred for 20 min. Quinine(8.92 g, 27.5 mmol) was added and the flask was degassed and backfilledwith nitrogen. The solution was cooled to −55° C. While stirring,methanol (3.04 mL, 75 mmol) was added. The mixture was stirred at −55°C. for 20 h. Upon warming to 25° C., the mixture was concentrated invacuo to a thick oil. The oil was dissolved in ethyl acetate (400 mL),washed with 1.0M aqueous hydrochloric acid solution (2×400 mL),saturated aqueous brine solution (100 mL), dried over magnesium sulfate,filtered, and concentrated in vacuo to afford the desired product,(1R,2S,3R,4S)-3-(methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (4.8 g, 24.5 mmol, 98%), as a clear waxy solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.26 (1H, d, J=8.5 Hz), 1.33 (1H, d, J=8.8 Hz), 3.00 (1H,s), 3.03 (1H, s), 3.21-3.30 (2H, m), 3.45 (3H, s), 6.02-6.04 (1H, m),6.14-6.16 (1H, m), 11.86 (1H, s).

b) Methyl(1S,2R,3S,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate

(1R,2S,3R,4S)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (4.61 g, 23.5 mmol) was dissolved in anhydrous tetrahydrofuran (40mL). The flask was degassed and backfilled with nitrogen and the mixturewas cooled to 0° C. Triethylamine (9.9 mL, 70.5 mmol) was added followedby the dropwise addition of ethyl chloroformate (4.48 mL, 47 mmol) withvigorous stirring. Immediate precipitation was observed. Additionaltetrahydrofuran (60 mL) was added. The mixture was stirred at 0° C. for1 h. Sodium azide (4.58 g, 70.5 mmol) was dissolved in water (30 mL) andadded to the reaction mixture at 0° C. The mixture was stirred at 0° C.for 5 min. The ice bath was removed. The mixture was warmed to 25° C.and was stirred for 2 h. The mixture was poured into water (300 mL) andthe product extracted into ethyl acetate (300 mL). The organic layer wasfurther washed with half-saturated aqueous sodium bicarbonate solution(2×100 mL), saturated aqueous brine solution (100 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. The oil was dissolved in anhydrous benzene (50 mL) and refluxedwhile stirring under nitrogen for 2 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a slightly yellow oil. Theoil was dissolved in dichloromethane (30 mL) and benzyl alcohol (2.68mL, 25.9 mmol) was added followed by triethylamine (6.61 mL, 47 mmol).The mixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C.the solution was concentrated in vacuo to afford a golden oil.Purification by flash column chromatography (Merck silica gel 60, 40-63μm, 15% ethyl acetate in hexanes) afforded the desired product, methyl(1S,2R,3S,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(5.51 g, 18.31 mmol, 78%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:1.38 (1H, d, J=9.1 Hz), 1.50 (1H, d, J=9.4 Hz), 3.10 (2H, s), 3.21 (1H,dd, J₁=9.2 Hz, J₂=2.3 Hz), 3.53 (3H, s), 4.62 (1H, dt, J₁=9.4 Hz, J₂=2.9Hz), 5.07 (2H, q, J=13.0 Hz), 5.29 (1H, d, J=8.6 Hz), 6.15-6.17 (1H, m),6.37-6.38 (1H, m), 7.29-7.35 (5H, m). LC-MS (ESI) calcd for C₁₇H₁₉NO₄301.13, found 258.1 (100%), 302.2 [M+H⁺] (70%), 603.5 [2M+H⁺] (20%).

c) Methyl (1R,2R,3S,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride

Methyl(1S,2R,3S,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(5.5 g, 18.27 mmol) was dissolved in ethyl acetate (75 mL). 5% Palladiumon carbon (650 mg) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 16 h.The mixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to afford a thick clear oil. The oil was dissolvedin ethyl acetate (15 mL) and added dropwise, with vigorous stirring, toa mixture of a 4.0M solution of hydrochloric acid in 1,4-dioxane (10 mL,40 mmol) in diethyl ether (90 mL). The desired product began toprecipitate as a white solid. The mixture was stirred for 20 min. Theprecipitate was collected by vacuum filtration, and was washed withadditional diethyl ether (15 mL). The solid was further dried in vacuofor 1 h to afford the desired product, methyl(1R,2R,3S,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylate hydrochloride(2.61 g, 12.69 mmol, 69%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.34-1.43 (4H, m), 1.54 (1H, d, J=9.5 Hz), 1.68 (1H, d, J=11.4 Hz),2.47-2.48 (2H, m), 3.03 (1H, dd, J=11.0 Hz, J₂=4.0 Hz), 3.49-3.53 (1H,m), 3.62 (3H, s), 8.07 (3H, bs). LC-MS (ESI) calcd for C₉H₁₅NO₂ (freeamine) 169.11, found 170.1 [M+H⁺] (100%), 339.2 [2M+H⁺] (50%).

d) Methyl(1R,2R,3S,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1R,2R,3S,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (1 g, 4.86 mmol) was dissolved in methanol (23 mL). Sodiumacetate (0.755 g, 9.2 mmol) was added followed by 4 Å powdered molecularsieves (1g) and 4-fluoro-benzaldehyde (0.571 g, 4.6 mmol). Sodiumcyanoborohydride (0.578 g, 9.2 mmol) was added and the mixture wasstirred at 25° C. for 16 h. The mixture was poured into a mixture ofsaturated aqueous sodium bicarbonate solution (200 mL) and ethyl acetate(300 mL). After shaking, both layers were passed through a plug ofCelite. The organic layer was further washed with saturated aqueoussodium bicarbonate solution (100 mL), saturated aqueous brine solution(100 mL), dried over magnesium sulfate, filtered, and concentrated invacuo to afford the crude product, methyl(1R,2R,3S,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(1.172 g, 4.23 mmol, 92%), as a clear oil. LC-MS (ESI) calcd forC₁₆H₂₀FNO₂ 277.15, found 278.2 [M+H⁺].

e)N-{3-[(1S,2S,7R,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1R,2R,3S,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.087 g, 0.3 mmol) was dissolved in anhydrous N,N-dimethylformamide(2.8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.3 mmol) was addedfollowed by N-methylmorpholine (0.07 mL, 0.63 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.061 g,0.315 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.126 mL, 0.9 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (25 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×25 mL), saturated aqueous brine solution (10 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. The oil was dissolved in methanol (4 mL) and the product wasprecipitated by the addition of 1.0M aqueous hydrochloric acid solution(4 mL) while stirring. The solid was collected by vacuum filtration andfurther dried in vacuo to afford the desired product,N-{3-[(1S,2S,7R,8R)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.0805 g, 0.144 mmol, 48%), as a white powder. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.23-1.48 (6H, m), 2.67-2.68 (2H, m), 3.06 (3H, s), 3.24(1H, d, J=15.0 Hz), 3.72 (1H, d, J=11.9 Hz), 4.07 (1H, d, J=15.6 Hz),5.12 (1H, d, J=15.7 Hz), 7.14 (2H, t, J=8.4 Hz), 7.39 (2H, dd, J₁=8.2Hz, J₂=5.8 Hz), 7.51 (1H, dd, J₁=8.4 Hz, J₂=2.3 Hz), 7.57-7.60 (2H, m),10.18 (1H, s). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₆S₂ 560.12, found 561.3[M+H⁺]. ee=99% [HPLC-analysis: Chiralpak AS-RH 4.6×250 mm, 5 micron atr.t., Solvent A-Solvent B (see table for gradient), 0.8 mL/min, 310 nm,t1=7.58 min (major), t2=10.08 min].

Example 5N-{3-[(1R,2R,7S,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid

The starting material (a) was prepared as described in J. Org. Chem.2000, 65, 6984-6991. cis-5-Norbornene-endo-2,3-dicarboxylic anhydride(8.21 g, 50 mmol) was suspended in a 1:1 mixture of toluene and carbontetrachloride (250 mL). The mixture was stirred for 10 min. Quinidine(17.84 g, 55 mmol) was added and the flask was degassed and backfilledwith nitrogen. The solution was cooled to −55° C. While stirring,methanol (6.08 mL, 150 mmol) was added. The mixture was stirred at −55°C. for 18 h. Upon warming to 25° C., the mixture was concentrated invacuo to a thick oil. The oil was dissolved in a mixture of ethylacetate (400 mL) and 1.0M aqueous hydrochloric acid solution (300 mL).After shaking, the layers were separated and the organic layer wasfurther washed with 1.0M aqueous hydrochloric acid solution (2×100 mL),saturated aqueous brine solution (100 mL), dried over magnesium sulfate,filtered, and concentrated in vacuo to afford the desired product,(1S,2R,3S,4R)-3-(methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (9.15 g, 46.6 mmol, 94%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.26 (1H, d, J=8.4 Hz), 1.33 (1H, d, J=8.4 Hz), 3.00 (1H, s), 3.03(1H, s), 3.21-3.29 (2H, m), 3.45 (3H, s), 6.02-6.04 (1H, m), 6.14-6.16(1H, m), 11.86 (1H, s).

b) Methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate

The intermediate (b) was prepared as described in Synthesis 2001, 11,1719-1730.(1S,2R,3S,4R)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (8.94 g, 45.57 mmol) was dissolved in anhydrous tetrahydrofuran(200 mL). The flask was degassed and backfilled with nitrogen and themixture was cooled to 0° C. Triethylamine (19.2 mL, 136.7 mmol) wasadded followed by the dropwise addition of ethyl chloroformate (8.69 mL,91.1 mmol) with vigorous stirring. Immediate precipitation was observed.The mixture was stirred at 0° C. for 1 h. Sodium azide (8.89 g, 136.7mmol) was dissolved in water (60 mL) and added to the reaction mixtureat 0° C. The mixture was stirred at 0° C. for 1 h. The ice bath wasremoved. The mixture was warmed to 25° C. and continued to stir for 2 h.The mixture was poured into water (400 mL) and the product extractedinto ethyl acetate (400 mL). The organic layer was further washed withhalf-saturated aqueous sodium bicarbonate solution (2×200 mL), saturatedaqueous brine solution (2×200 mL), dried over magnesium sulfate,filtered, and concentrated in vacuo to afford a slightly brown oil. Theoil was dissolved in anhydrous benzene (100 mL) and refluxed whilestirring under nitrogen for 2 h. Upon cooling to 25° C. the solution wasconcentrated in vacuo to afford a slightly brown oil. The oil wasdissolved in dichloromethane (60 mL) and benzyl alcohol (5.19 mL, 50.13mmol) was added followed by triethylamine (12.81 mL, 91.14 mmol). Themixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a golden oil. Purificationby flash column chromatography (Merck silica gel 60, 40-63 μm, 10% ethylacetate in hexanes) afforded the desired product, methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(10.1 g, 33.55 mmol, 74%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:1.38 (1H, d, J=8.7 Hz), 1.50 (1H, d, J=8.4 Hz), 3.10 (2H, s), 3.21 (1H,d, J=8.8 Hz), 3.53 (3H, s), 4.59-4.64 (1H, m), 5.07 (2H, q, J=13.0 Hz),5.29 (1H, d, J=8.3 Hz), 6.15-6.17 (1H, m), 6.37-6.38 (1H, m), 7.27-7.36(5H, m). LC-MS (ESI) calcd for C₁₇H₁₉NO₄ 301.13, found 258.1 (100%),302.2 [M+H⁺] (70%), 603.5 [2M+H⁺] (20%).

c) Methyl (1S,2S,3R,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride

Methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(10 g, 33.22 mmol) was dissolved in ethyl acetate (150 mL). 5% Palladiumon carbon (1.5 mg) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 2 h. Themixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to a volume of 50 mL. The solution was addeddropwise, with vigorous stirring, to a mixture of 4.0 M hydrochloricacid solution in 1,4-dioxane (20 mL) in diethyl ether (200 mL). Thedesired product began to precipitate as a white solid. The mixture wasstirred for 10 min. The precipitate was collected by vacuum filtration,washed with additional diethyl ether (15 mL). The solid was furtherdried in vacuo for 1 h to afford the desired product, methyl(1S,2S,3R,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylate hydrochloride(5.21 g, 25.33 mmol, 76.3%), as a white powder. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.33-1.42 (4H, m), 1.54 (1H, d, J=10.3 Hz), 1.69 (1H, d,J=11.5 Hz), 2.46-2.48 (2H, m), 3.03 (1H, dd, J₁=10.8 Hz, J₂=4.1 Hz),3.46-3.55 (1H, m), 3.62 (3H, s), 8.09 (3H, bs). LC-MS (ESI) calcd forC₉H₁₅NO₂ (free amine) 169.11, found 170.1 [M+H⁺] (100%), 339.2 [2M+H⁺](50%).

d) Methyl(1S,2S,3R,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1S,2S,3R,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (1 g, 4.86 mmol) was dissolved in methanol (23 mL). Sodiumacetate (0.755 g, 9.2 mmol) was added followed by 4 Å powdered molecularsieves (1g) and 4-fluoro-benzaldehyde (0.571 g, 4.6 mmol). Sodiumcyanoborohydride (0.578 g, 9.2 mmol) was added and the mixture wasstirred at 25° C. for 16 h. The mixture was poured into a mixture ofsaturated aqueous sodium bicarbonate solution (200 mL) and ethyl acetate(300 mL). After shaking, both layers were passed through a plug ofCelite. The organic layer was further washed with saturated aqueoussodium bicarbonate solution (100 mL), saturated aqueous brine solution(100 mL), dried over magnesium sulfate, filtered, and concentrated invacuo to afford the crude product, methyl(1S,2S,3R,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(1.11 g, 4.0 mmol, 87%), as a clear oil. LC-MS (ESI) calcd forC₁₆H₂₀FNO₂ 277.15, found 278.2 [M+H⁺].

e)N-{3-[(1R,2R,7S,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1S,2S,3R,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.087 g, 0.3 mmol) was dissolved in anhydrous N,N-dimethylformamide(2.8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.3 mmol) was addedfollowed by N-methylmorpholine (0.07 mL, 0.63 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.061 g,0.315 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.126 mL, 0.9 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (25 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×25 mL), saturated aqueous brine solution (10 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. The oil was dissolved in methanol (4 mL) and the product wasprecipitated by the addition of 1.0M aqueous hydrochloric acid solution(4 mL) while stirring. The solid was collected by vacuum filtration andfurther dried in vacuo to afford the desired product,N-{3-[(1R,2R,7S,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.0781 g, 0.139 mmol, 46%), as a white powder. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.23-1.48 (6H, m), 2.67-2.68 (2H, m), 3.06 (3H, s), 3.24(1H, d, J=15.0 Hz), 3.72 (1H, d, J=11.9 Hz), 4.07 (1H, d, J=15.6 Hz),5.12 (1H, d, J=15.7 Hz), 7.14 (2H, t, J=8.4 Hz), 7.39 (2H, dd, J₁=8.2Hz, J₂=5.8 Hz), 7.51 (1H, dd, J₁=8.4 Hz, J₂=2.3 Hz), 7.57-7.60 (2H, m),10.18 (1H, s). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₆S₂ 560.12, found 561.3[M+H⁺]. ee=99% [HPLC-analysis: Chiralpak AS-RH 4.6×250 mm, 5 micron atr.t., Solvent A-Solvent B (see table for gradient), 0.8 mL/min, 310 nm,t1=7.58 min, t2=10.08 min (major)].

Example 6N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2S,3R,4R)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid

The starting material (a) was prepared as described in J. Org. Chem.2000, 65, 6984-6991. cis-5-Norbornene-exo-2,3-dicarboxylic anhydride (5g, 30.45 mmol) was suspended in a 1:1 mixture of toluene and carbontetrachloride (610 mL). The mixture was stirred for 10 min. Quinine(10.87 g, 33.5 mmol) was added and the flask was degassed and backfilledwith nitrogen. The solution was cooled to −55° C. While stirring,methanol (3.7 mL, 91.35 mmol) was added. The mixture was stirred at −55°C. for 16 h. Upon warming to 25° C., the mixture was concentrated invacuo to a foam. The foam was dissolved in a mixture of ethyl acetate(400 mL) and 1.0M aqueous hydrochloric acid solution (400 mL). Thelayers were separated and the organic layer was further washed with 1.0Maqueous hydrochloric acid solution (2×200 mL), saturated aqueous brinesolution (100 mL) and dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the desired product,(1S,2S,3R,4R)-3-(methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (5.95 g, 30.3 mmol, 99%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.31 (1H, d, J=8.5 Hz), 1.98 (1H, d, J=8.6 Hz), 2.51 (2H, d, J=1.6Hz), 2.95 (2H, bs), 3.52 (3H, s), 6.17-6.21 (2H, m), 12.16 (1H, s).

b) Methyl(1R,2R,3S,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate

(1S,2S,3R,4R)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (5.9 g, 30 mmol) was dissolved in anhydrous tetrahydrofuran (133mL). The flask was degassed and backfilled with nitrogen and the mixturewas cooled to 0° C. Triethylamine (12.64 mL, 90 mmol) was added followedby the dropwise addition of ethyl chloroformate (5.72 mL, 60 mmol) withvigorous stirring. Immediate precipitation was observed. The mixture wasstirred at 0° C. for 1 h. Sodium azide (5.86 g, 90 mmol) was dissolvedin water (40 mL) and added to the reaction mixture at 0° C. The mixturewas stirred at 0° C. for 5 min. The ice bath was removed. The mixturewas warmed to 25° C. and continued to stir for 2 h. The mixture waspoured into water (300 mL) and the product extracted into ethyl acetate(300 mL). The organic layer was further washed with half-saturatedaqueous sodium bicarbonate solution (2×100 mL), saturated aqueous brinesolution (100 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford a light brown oil. The oil was dissolvedin anhydrous benzene (66 mL) and refluxed while stirring under nitrogenfor 2 h. Upon cooling to 25° C. the solution was concentrated in vacuoto afford a light brown oil. The oil was dissolved in dichloromethane(40 mL) and benzyl alcohol (3.41 mL, 33 mmol) was added followed bytriethylamine (8.44 mL, 60 mmol). The mixture was refluxed undernitrogen for 16 h. Upon cooling to 25° C. the solution was concentratedin vacuo to afford a thick oil. Purification by flash columnchromatography (Merck silica gel 60, 40-63 μm; 1^(st) column: 3:1hexanes/ethyl acetate; 2^(nd) column: 2:4:1dichloromethane/pentane/diethyl ether) afforded the desired product,methyl(1R,2R,3S,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(6.95 g, 23.09 mmol, 77%), as a pale yellow oil. ¹H NMR (400 MHz, CDCl₃)δ: 1.59 (1H, d, J=9.3 Hz), 1.96 (1H, d, J=9.3 Hz), 2.66 (1H, d, J=7.9Hz), 2.75 (1H, s), 2.96 (1H, s), 3.59 (3H, s), 4.01 (1H, t, J=8.5 Hz),5.09 (2H, q, J=10.4 Hz), 5.46 (1H, d, J=9.4 Hz), 6.17-6.22 (2H, m),7.29-7.36 (5H, m). LC-MS (ESI) calcd for C₁₇H₁₉NO₄ 301.13, found 258.1(100%), 302.2 [M+H⁺] (70%), 603.5 [2M+H⁺] (20%).

c) Methyl (1S,2R,3S,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride

Methyl(1R,2R,3S,4S)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(1 g, 3.32 mmol) was dissolved in ethyl acetate (15 mL). 5% Palladium oncarbon (120 mg) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 16 h.The mixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to afford a thick clear oil. The oil was dissolvedin diethyl ether (10 mL) and added dropwise, with vigorous stirring, toa mixture of 4.0M hydrochloric acid solution in 1,4-dioxane (1.8 mL) indiethyl ether (18 mL). The desired product began to precipitate as awhite solid. Additional diethyl ether (10 mL) was added and the mixturewas stirred for 10 min. The precipitate was collected by vacuumfiltration, washed with additional diethyl ether (2×8 mL). The solid wasfurther dried in vacuo for 1 h to afford the desired product, methyl(1S,2R,3S,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylate hydrochloride(0.64 g, 3.11 mmol, 94%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.17-1.27 (3H, m), 1.40-1.61 (2H, m), 1.91 (1H, d, J=10.7 Hz), 2.36(1H, d, J=4.1 Hz), 2.44 (1H, d, J=3.1 Hz), 2.75 (1H, d, J=7.8 Hz),3.30-3.38 (1H, m), 3.61 (3H, s), 8.05 (3H, bs). LC-MS (ESI) calcd forC₉H₁₅NO₂ (free amine) 169.11, found 170.3 [M+H⁺] (100%), 339.3 [2M+H⁺](50%).

d) Methyl(1S,2R,3S,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1S,2R,3S,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (prepared as described in Example 6c, 0.5 g, 2.43 mmol)was dissolved in methanol (12 mL). Sodium acetate (0.4 g, 4.86 mmol) wasadded followed by 4 Å powdered molecular sieves (0.5 g) and4-fluoro-benzaldehyde (0.302 g, 2.43 mmol). Sodium cyanoborohydride(0.305 g, 4.86 mmol) was added and the mixture was stirred at 25° C. for16 h. The mixture was poured into a mixture of saturated aqueous sodiumbicarbonate solution (200 mL) and ethyl acetate (300 mL). After shaking,both layers were passed through a plug of Celite. The organic layer wasfurther washed with saturated aqueous sodium bicarbonate solution (100mL), saturated aqueous brine solution (100 mL), dried over magnesiumsulfate, filtered, and concentrated in vacuo to afford the crudeproduct, methyl(1S,2R,3S,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.663 g, 2.39 mmol, 98%), as a clear oil. LC-MS (ESI) calcd forC₁₆H₂₀FNO₂ 277.15, found 278.2 [M+H⁺].

e)N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1S,2R,3S,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.6 g, 2.16 mmol) was dissolved in anhydrous N,N-dimethylformamide (20mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.72 g, 2.16 mmol) was addedfollowed by N-methylmorpholine (0.5 mL, 4.54 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.435 g,2.27 mmol) was added and the mixture was stirred at 25° C. for 45 min.Triethylamine (0.91 mL, 6.48 mmol) was added and the mixture was stirredat 50° C. for 16 h.

Upon cooling to 25° C., the solution was diluted with ethyl acetate (300mL) and washed with 1.0M aqueous hydrochloric acid solution (3×300 mL),saturated aqueous brine solution (100 mL), dried over magnesium sulfate,filtered, and concentrated in vacuo to afford a golden oil. Purificationby flash column chromatography (Merck silica gel 60, 40-63 μm, 0 to0.75% methanol in dichloromethane) afforded the product as white foam.The foam was dissolved in methanol (10 mL) and the product wasprecipitated by the addition of a 1.0M aqueous hydrochloric acidsolution (20 mL) while stirring. The solid was collected by vacuumfiltration and further dried in vacuo to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.573 g, 1.02 mmol, 47%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.16-1.22 (2H, m), 1.37-1.65 (4H, m), 2.49-2.53 (1H, m), 2.63 (1H, d,J=2.3 Hz), 3.02 (1H, d, J=8.5 Hz), 3.05 (3H, s), 3.52 (1H, d, J=9.4 Hz),4.41 (1H, d, J=15.6 Hz), 4.95 (1H, d, J=15.6 Hz), 7.14 (2H, t, J=9.0Hz), 7.32 (2H, dd, J₁=8.1 Hz, J₂=5.7 Hz), 7.50 (1H, dd, J₁=9.5 Hz,J₂=2.3 Hz), 7.55-7.57 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₅H₂₅FN₄O₆S₂ 560.12, found 561.3 [M+H⁺]. ee=90% [HPLC-analysis:Chiralpak AS-RH 2.1×150 mm, 5 micron at r.t., Solvent A-Solvent B (seetable for gradient), 0.3 mL/min, 312 nm, t1=4.3 min (major), t2=6.0min].

Alternatively,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamidecan be prepared as follows:

f) (rac-di-exo)-3-Aza-tricyclo[4.2.1.0^(2,5)]nonan-4-one

Bicyclo[2.2.1]hept-2-ene (1000 g, 10.6 mol) was dissolved in ethylacetate (1.7 L) and the resulting solution was cooled to 0° C.Chlorosulfonyl isocyanate (969 mL, 11.1 mol) was added at 0-20° C. over30 min. The mixture was allowed to warm to 25° C. and stirred for 4 h,then cooled to 0° C. A mixture of sodium sulfite (1500 g, 11.9 mol) inwater (6 L) was added at 0-20° C. The milky suspension was stirred at25° C. for 30 min and cooled to 0° C. A 50% queous sodium hydroxidesolution (1.6 L, 30.3 mol) was added at 0-15° C. to adjust to pH 7. Asaturated aqueous sodium carbonate solution (300 mL) was added to adjustthe pH to 7.5-8.0. The mixture was filtered and the solid was washedwith ethyl acetate (3×2 L) and the solid was discarded. The combinedethyl acetate extracts were washed with saturated aqueous brine solution(2 L), dried over magnesium sulfate and filtered. The solution wasconcentrated in vacuo to dryness to afford the desired product,(rac-di-exo)-3-aza-tricyclo[4.2.1.0^(2,5)]nonan-4-one (1220 g, 8.9 mol,84%), as a white glassy solid. ¹H NMR (400 MHz, CDCl₃) δ 1.02-1.11 (2H,m), 1.24 (1H, dt, J₁=10.9 Hz, J₂=1.6 Hz), 1.51-1.72 (3H, m), 2.37-2.37(1H, m), 2.43-2.44 (1H, m), 2.99-3.00 (1H, m), 3.40 (1H, d, J=3.4 Hz),5.73 (1H, bs).

g) (rac-di-exo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acidhydrochloride

To (rac-di-exo)-3-aza-tricyclo[4.2.1.0^(2,5)]nonan-4-one (23.37 g, 170.4mmol) was added a 12.0M aqueous hydrochloric acid solution (150 mL). Themixture was stirred at 25° C. for 12 h. The solvent was evaporated invacuo and the crude compound was dried under high vacuum for 0.5 h. Thecrude compound was triturated with acetone and filtered to afford(rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acidhydrochloride (28.43 g, 148.3 mmol, 87%), as a white solid. ¹H NMR (400MHz, DMSO-d₆) δ 1.15-1.26 (3H, m), 1.42-1.59 (2H, m), 1.87 (1H, d,J=10.3 Hz), 2.33 (1H, d, J=3.4 Hz), 2.45 (1H, d, J=2.3 Hz), 2.67 (1H, d,J=7.6 Hz), 3.23-3.26 (1H, m), 7.93 (3H, bs), 12.73 (1H, bs).

h) (rac-di-exo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester hydrochloride

To absolute ethanol (75 mL) at −10° C. was added thionyl chloride (4.1mL, 54.5 mmol) dropwise followed by(rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acidhydrochloride (9.60 g, 50.1 mmol). The mixture was stirred at 0° C. for1 h, at 25° C. for 4 h, and heated at reflux for 0.5 h. The solution wasconcentrated in vacuo and dried under high vacuum to afford the crude(rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethyl esterhydrochloride (11.01 g, 50.1 mmol, 100%), as an off-white solid. ¹H NMR(400 MHz, DMSO-d₆) δ 1.17-1.27 (3H, m), 1.21 (3H, t, J=7.0 Hz),1.43-1.57 (2H, m), 1.91 (1H, d, J=10.0 Hz), 2.36 (1H, d, J=3.9 Hz), 2.42(1H, d, J=3.0 Hz), 2.72 (1H, d, J=7.6 Hz), 3.28 (1H, d, J=8.3 Hz),4.00-4.13 (2H, m), 8.06 (3H, bs).

i) (rac-di-exo)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester

To (rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester hydrochloride (11.01 g, 50.1 mmol) was added saturated aqueoussodium bicarbonate solution (50 mL) and the mixture was stirred at 25°C. for 0.5 h. The crude product was extracted with ethyl acetate (3×100mL). The solution was dried over magnesium sulfate, filtered, andconcentrated in vacuo and dried under high vacuum for 2 h to afford thecrude (rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (8.17 g, 44.6 mmol, 89%), as a brown oil. ¹H NMR (400 MHz, CDCl₃)δ 1.10-1.26 (3H, m), 1.29 (3H, t, J=7.0 Hz), 1.45-1.62 (2H, m), 1.86(2H, bs), 1.95 (1H, dt, J₁=10.3 Hz, J₂=1.9 Hz), 2.09 (1H, d, J=4.5 Hz),2.49 (1H, d, J=4.2 Hz), 2.56 (1H, d, J=9.0 Hz), 3.24 (1H, d, J=7.7 Hz),4.09-4.21 (2H, m).

j) (1R,2S,3R,4S)-3-Ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate

To a solution of (rac-di-exo)-3-amino-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (408.47 g, 2.98 mol) in ethyl acetate (500 mL) wasadded a solution of (1S)-(+)-10-camphorsulfonic acid (691.70 g, 2.98mol) in ethanol (800 mL) at 50-75° C. over 30 min. The resultingsolution was stirred at 70° C. for 1 h. More ethyl acetate (2.7 L) wasadded at >55° C. The solution was allowed to cool to 50° C. and seededwith (1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (ca. 20 mg). The mixture was allowed tocool to 25° C. and stirred for 16 h. The suspension was filtered and thewet filter cake was washed with ethyl acetate (2×500 mL). The crude saltwas recrystallized from ethanol (600 mL) and ethyl acetate (3 L) toafford the desired product,(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (334.84 g, 0.806 mol, 27%, >99.5% de), asa white solid. ¹H NMR (400 MHz, CDCl₃) δ 0.84 (3H, s), 1.08 (3H, s),1.30 (3H, t, J=6.9 Hz), 1.32-1.43 (4H, m), 1.58-1.75 (3H, m), 1.89 (1H,d, J=17.7 Hz), 1.95-2.07 (3H, m), 2.33 (1H, dt, J₁=18.4 Hz, J₂=3.9 Hz),2.53 (1H, s), 2.58-2.65 (1H, m), 2.69 (1H, d, J=2.9 Hz), 2.76-2.79 (2H,m), 3.26 (1H, d, J=14.1 Hz), 3.60 (1H, d, J=7.4 Hz), 4.14-4.27 (2H, m),7.80 (3H, bs).

Alternatively,(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate can be prepared as follows:

(rac-di-exo)-3-Aza-tricyclo[4.2.1.0^(2,5)]nonan-4-one (prepared asdescribed in Example 6f, 1220 g, 8.9 mol) was dissolved in ethyl acetate(1.7 L). The solution was heated to 50° C. and a solution of(1S)-(+)-10-camphorsulfonic acid (2066 g, 8.9 mol) in ethanol (2.5 L) at50-75° C. over 30 min. The resulting solution was stirred at 70° C. for2 h. More ethyl acetate (8 L) was added causing the temperature to dropto >55° C. and the solution was seeded with(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (ca. 100 mg). The mixture was allowed tocool to 25° C. and stirred for 16 h. The precipitate was collected byfiltration and the wet filter cake was washed with ethyl acetate (2×2L). The crude salt was dried at 25° C. for 48 h and then wasrecrystallized from ethanol (2 L) and ethyl acetate (2.5 L) to affordthe desired product,(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (920 g, 2.21 mol, 25%, >99.9% de), as awhite solid.

k) (1S,2R,3S,4R)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester

To (1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (2.76 g, 6.64 mmol) was added ethylacetate (28 mL) and saturated aqueous sodium carbonate solution (28 mL)and the mixture was stirred at 25° C. for 0.5 h. The organic layer wasseparated and the aqueous layer was extracted with ethyl acetate (2×50mL). The solution was dried over magnesium sulfate, filtered, andconcentrated in vacuo and dried under high vacuum for 1 h to afford(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (1.15 g, 6.28 mmol, 95%), as a colorless oil. ¹H NMR (400 MHz,CDCl₃) δ 1.10-1.26 (3H, m), 1.29 (3H, t, J=7.0 Hz), 1.45-1.62 (2H, m),1.86 (2H, bs), 1.95 (1H, dt, J₁=10.3 Hz, J₂=1.9 Hz), 2.09 (1H, d, J=4.5Hz), 2.49 (1H, d, J=4.2 Hz), 2.56 (1H, d, J=9.0 Hz), 3.24 (1H, d, J=7.7Hz), 4.09-4.21 (2H, m).

In order to determine the enantiomeric excess,(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester was derivatized to the (S)-mandelate salt as follows: To asolution of (1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (34.2 mg, 0.187 mmol) in ethyl acetate (1 mL) was added(S)-α-hydroxyphenylacetic acid (28.7 mg, 0.187 mmol) and the mixture wasstirred at 25° C. for 0.5 h. The solid was filtered and dried under highvacuum to afford(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(S)-α-hydroxyphenylacetate (11.4 mg, 0.034 mmol, 18%, de=97%), as awhite solid. ¹H NMR (400 MHz, CDCl₃) δ 1.08-1.20 (3H, m), 1.28 (3H, t,J=7.1 Hz), 1.50-1.59 (2H, m), 1.79 (1H, d, J=10.9 Hz), 2.23 (1H, s),2.46-2.48 (2H, m), 3.04 (1H, d, J=7.8 Hz), 4.05-4.18 (2H, m), 4.89 (1H,s), 5.49 (3H, bs), 7.22-7.31 (3H, m), 7.43 (2H, d, J=6.9 Hz).

l)(1S,2R,3S,4R)-3-(4-Fluorobenzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

To a solution of(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (1.15 g, 6.28 mmol) in ethanol (30 mL) was added4-fluorobenzaldehyde (0.68 mL, 6.31 mmol), glacial acetic acid (0.4 mL,6.99 mmol), and sodium cyanoborohydride (1.04 g, 15.7 mmol) at 25° C.After stirring for 3 h, the mixture was diluted with ethyl acetate (50mL) and quenched with saturated aqueous sodium bicarbonate solution (50mL) for 0.5 h. The mixture was filtered through Celite. The organiclayer was separated and the aqueous layer was extracted with ethylacetate (2×50 mL). When all solvent was removed, a solid was formed. Thesolid was filtered, washed with water, and dried in vacuo to afford thedesired product,(1S,2R,3S,4R)-3-(4-fluorobenzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (1.74 g, 5.97 mmol, 95%), as a white solid. ¹H NMR (400MHz, CDCl₃) δ 1.05-1.16 (2H, m), 1.21 (1H, dt, J₁=8.0 Hz, J₂=1.6 Hz),1.27 (3H, t, J=7.4 Hz), 1.45-1.61 (2H, m), 1.94 (1H, dt, J₁=10.1 Hz,J₂=1.9 Hz), 2.28 (1H, d, J=3.9 Hz), 2.43 (1H, d, J=3.3 Hz), 2.60 (1H,dd, J₁=8.8 Hz, J₂=1.5 Hz), 2.94 (1H, d, J=7.8 Hz), 3.66 (1H, d, J=13.2Hz), 3.80 (1H, d, J=13.5 Hz), 4.13 (2H, q, J=7.0 Hz), 6.97 (2H, t, J=8.5Hz), 7.26 (2H, t, J=7.1 Hz).

Alternatively,(1S,2R,3S,4R)-3-(4-Fluorobenzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester can be prepared as follows:

(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (prepared as described in Example 6j, 2000g, 4.81 mol) and powdered potassium carbonate (1320 g, 9.62 mol) weresuspended in ethyl acetate (20 L). The suspension was stirred at 25° C.for 16 h and filtered. The ethyl acetate filtrate was concentrated invacuo to afford the free amine (1050 g) as a liquid. The liquid wasdissolved in ethanol (10 L), and 4-fluorobenzaldehyde (558 mL, 5.3 mol)and acetic acid (362 mL, 6.3 mol) were added, causing the temperature torise to 28-30° C. The solution was allowed to cool to 25° C. and stirredfor 30 min. A cloudy solution of sodium cyanoborohydride (756 g, 12.03mol) in ethanol (5 L) was added in 20 min, causing the temperature torise to 45-50° C. The mixture was allowed to cool to 25° C. and stirredfor 16 h. The mixture was concentrated in vacuo to a volume of about13-14 L. Water (1-2 L) was added, and the resulting mixture was furtherconcentrated in vacuo. A saturated aqueous sodium bicarbonate solution(4 L) and water (4 L) were added with stirring. The pH was adjusted to8.0-8.5 by adding additional saturated aqueous sodium bicarbonatesolution (˜500 mL). The mixture was stirred for 1 h before the solidswere collected by filtration and the wet filter cake was washed withwater (2 L). The solid was dried in vacuo at 35° C. for 64 h to affordthe desired product,(1S,2R,3S,4R)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (1350 g, 4.63 mol, 96%), as a white solid.

m)(1S,2R,3S,4R)-3-{(4-Fluorobenzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

To a solution of(1S,2R,3S,4R)-3-(4-fluorobenzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (100.6 mg, 0.345 mmol) in N,N-dimethylformamide (3.0mL) was added(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 120.8 mg, 0.362 mmol),4-dimethylaminopyridine (10.6 mg, 0.086 mmol), and1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (70.9 mg,0.362 mmol). After stirring at 25° C. for 12 h, the mixture was dilutedwith ethyl acetate and acidified with 1.0M aqueous hydrochloric acidsolution to pH 1. The organic layer was separated and the aqueous layerwas extracted with ethyl acetate (2×20 mL). The combined organic layerwas dried over magnesium sulfate, filtered, and concentrated in vacuo,and dried under high vacuum to afford the crude product,(1S,2R,3S,4R)-3-{(4-fluorobenzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as a faintly yellow oil. The crude product was used inthe next step without further purification. LC-MS (ESI) calcd forC₂₇H₃₁FN₄O₇S₂ 606.16, found 607.2 [M+H⁺].

n)N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a solution of the crude(1S,2R,3S,4R)-3-{(4-fluorobenzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester in absolute ethanol (3 mL) was added a 21 wt. %solution of sodium ethoxide in ethanol (0.51 mL, 1.37 mmol). Afterstirring at 60° C. for 2 h, the mixture was diluted with ethyl acetateand acidified with 1.0M aqueous hydrochloric acid solution to pH 1. Theorganic layer was separated and the aqueous layer was extracted withethyl acetate (2×20 mL). The combined organic layer was dried overmagnesium sulfate, filtered, and concentrated in vacuo. The crudemixture was purified by flash column chromatography (Teledyne IscoRediSep column; 0 to 100% ethyl acetate in hexanes) to afford thedesired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(131.5 mg, 0.235 mmol, 68% over two steps), as an off-white solid. ¹HNMR (400 MHz, CD₃OD) δ 1.28 (2H, d, J=11.0 Hz), 1.47 (1H, t, J=10.8 Hz),1.57-1.74 (3H, m), 2.56 (1H, d, J=3.2 Hz), 2.75 (1H, d, J=2.3 Hz), 2.96(1H, d, J=9.2 Hz), 3.02 (3H, s), 3.58 (1H, d, J=9.2 Hz), 4.42 (1H, d,J=15.5 Hz), 5.03 (1H, d, J=15.7 Hz), 7.04 (2H, t, J=8.5 Hz), 7.31 (2H,dd, J₁=7.9 Hz, J₂=5.5 Hz), 7.37 (1H, d, J=8.8 Hz), 7.54 (1H, dd, J₁=8.3Hz, J₂=2.3 Hz), 7.69 (1H, d, J=2.3 Hz). LC-MS (ESI) calcd forC₂₅H₂₅FN₄O₆S₂ 560.12, found 561.4 [M+H⁺]. ee=98.5% [HPLC-analysis:Chiralpak AS-RH 2.1×150 mm, 5 micron at r.t., Solvent A-Solvent B (seetable for gradient), 0.3 mL/min, 312 nm, t1=7.58 min (major), t2=8.95min].

Alternatively,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamidecan be prepared as follows:

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 1.88 kg, 5.63 mol) and(1S,2R,3S,4R)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 6l, 1.72 kg, 5.91mol) were dissolved in acetonitrile (18.8 L) at 23° C.N-Methylmorpholine (1.25 kg, 12.4 mol) was added and the resultingsuspension was stirred at 23° C. for 1 h. The suspension was cooled to0° C. and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride(1.19 kg, 6.20 mol) was added in one portion. The mixture was stirred at0° C. for 3 h, and was then allowed to warm to 23° C. and stirredovernight. Triethylamine (1.88 kg, 18.6 mol) was added and the mixturewas then heated at 50° C. for 3 h. The mixture was partiallyconcentrated in vacuo at 45° C., and was then diluted with ethyl acetate(22.5 L) and washed with 2.0M aqueous hydrochloric acid solution (22.6L). The resulting aqueous fraction was extracted with ethyl acetate(2×9.4 L). The combined organic extracts were washed with 1.0M aqueoushydrochloric acid solution (10.4 L) and then with water (18.8 L). Theresulting organic fraction was filtered through Celite (600 g), and thefiltrate was then partially concentrated in vacuo at 45° C. Absoluteethanol (5.6 L) was added to the residue, and the mixture was thenheated at 50° C. with stirring. Dichloromethane (400 mL) was added inportions until crystallization initiated. Absolute ethanol (20.7 L) wasadded in portions over 1 h, and the resulting mixture was stirred at 23°C. overnight. The mixture was filtered and the solid was then washedwith absolute ethanol (1.9 L). The solid was further dried in vacuo at45° C. to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(2.46 kg, 4.39 mol, 78%), as an off-white crystalline solid.

X-ray data—FIG. 1 shows a x-ray diffraction ofN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(as prepared in Example 6 on a kg scale). In the x-ray graph, the angleof diffraction 2 theta is plotted on the x-axis and the peak intensityis plotted on the y-axis. The strongest lines in the x-ray diffractiongraph are observed at angles of 6.2°, 17.9°, 19.7°, 20.5°, 22.6°, and24.8°±0.3°, with lesser intensity lines at 12.4°, 16.5°, 18.7°, 21.6°,23.2°, 24.1°, 25.6°, 26.6°, 27.1°, 28.5°, and 29.3°.

IR data—FIG. 2 shows a FT-Raman spectrum ofN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(as prepared in Example 6 on a kg scale), which is characterized by thefollowing major IR bands at 1617, 1524, 1321, 1260, 1229, 1217, and 1163cm⁻¹, with minor bands at 1498, 1465, 1147, 836, 727, and 406 cm⁻¹.

Example 7N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1R,2R,3S,4S)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid

The starting material (a) was prepared as described in J. Org. Chem.2000, 65, 6984-6991. cis-5-Norbornene-exo-2,3-dicarboxylic anhydride (5g, 30.45 mmol) was suspended in a 1:1 mixture of toluene and carbontetrachloride (150 mL). The mixture was stirred for 10 min. Quinidine(10.9 g, 33.5 mmol) was added and the flask was degassed and backfilledwith nitrogen. The solution was cooled to −55° C. While stirring,methanol (3.7 mL, 91.35 mmol) was added. The mixture was stirred at −55°C. for 16 h. Upon warming to 25° C., the mixture was concentrated invacuo to a foam. The foam was dissolved in a mixture of ethyl acetate(400 mL) and 1.0M aqueous hydrochloric acid solution (400 mL). Thelayers were separated and the organic layer was further washed with 1.0Maqueous hydrochloric acid solution (2×100 mL), saturated aqueous brinesolution (100 mL) and dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the desired product,(1R,2R,3S,4S)-3-(methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (5.92 g, 30.2 mmol, 99%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.29 (1H, d, J=10.2 Hz), 1.96 (1H, d, J=8.6 Hz), 2.47-2.49 (2H, m),2.93-2.94 (2H, m), 3.51 (3H, s), 6.15-6.20 (2H, m), 12.15 (1H, s).

b) Methyl(1S,2S,3R,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate

The intermediate (b) was prepared as described in Synthesis 2001, 11,1719-1730.(1R,2R,3S,4S)-3-(Methoxycarbonyl)bicyclo[2.2.1]hept-5-ene-2-carboxylicacid (5.9 g, 30 mmol) was dissolved in anhydrous tetrahydrofuran (133mL). The flask was degassed and backfilled with nitrogen and the mixturewas cooled to 0° C. Triethylamine (12.64 mL, 90 mmol) was added followedby the dropwise addition of ethyl chloroformate (5.72 mL, 60 mmol) withvigorous stirring. Immediate precipitation was observed. The mixture wasstirred at 0° C. for 1 h. Sodium azide (5.86 g, 90 mmol) was dissolvedin water (40 mL) and added to the reaction mixture at 0° C. The mixturewas stirred at 0° C. for 5 min. The ice bath was removed. The mixturewas warmed to 25° C. and continued to stir for 2 h. The mixture waspoured into water (300 mL) and the product extracted into ethyl acetate(350 mL). The organic layer was further washed with half-saturatedaqueous sodium bicarbonate solution (2×100 mL), saturated aqueous brinesolution (100 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford a light brown oil.

The oil was dissolved in anhydrous benzene (66 mL) and refluxed whilestirring under nitrogen for 2 h. Upon cooling to 25° C. the solution wasconcentrated in vacuo to afford a light yellow oil. The oil wasdissolved in dichloromethane (40 mL) and benzyl alcohol (3.41 mL, 33mmol) was added followed by triethylamine (8.44 mL, 60 mmol). Themixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a thick oil. Purificationby flash column chromatography (Merck silica gel 60, 40-63 μm; 1^(st)column: 3:1 hexanes/ethyl acetate; 2nd column: 2:4:1dichloromethane/pentane/diethyl ether) afforded the desired product,methyl(1S,2S,3R,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(6.195 g, 20.58 mmol, 69%), as a faintly yellow oil. ¹H NMR (400 MHz,CDCl₃) δ: 1.60 (1H, d, J=9.4 Hz), 1.97 (1H, d, J=9.3 Hz), 2.66 (1H, d,J=7.5 Hz), 2.75 (1H, s), 2.96 (1H, s), 3.60 (3H, s), 4.02 (1H, t, J=8.9Hz), 5.09 (2H, q, J=10.5 Hz), 5.47 (1H, d, J=8.8 Hz), 6.18-6.23 (2H, m),7.29-7.37 (5H, m). LC-MS (ESI) calcd for C₁₇H₁₉NO₄ 301.13, found 258.1(100%), 302.2 [M+H⁺] (70%), 603.4 [2M+H⁺] (20%).

c) Methyl (1R,2S,3R,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride

Methyl(1S,2S,3R,4R)-3-{[(benzyloxy)carbonyl]amino}bicyclo[2.2.1]hept-5-ene-2-carboxylate(1 g, 3.32 mmol) was dissolved in ethyl acetate (15 mL). 5% Palladium oncarbon (120 mg) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 16 h.The mixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to afford a thick clear oil. The oil was dissolvedin diethyl ether (10 mL) and added dropwise, with vigorous stirring, toa mixture of 4.0M hydrochloric acid solution in 1,4-dioxane (1.8 mL, 7.2mmol) in diethyl ether (18 mL). The desired product began to precipitateas a white solid. Additional diethyl ether (10 mL) was added and themixture was stirred for 10 min. The precipitate was collected by vacuumfiltration and washed with additional diethyl ether (2×8 mL). The solidwas further dried in vacuo for 1 h to afford the desired product, methyl(1R,2S,3R,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylate hydrochloride(0.554 g, 2.7 mmol, 81%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.18-1.27 (3H, m), 1.37-1.61 (2H, m), 1.90 (1H, d, J=11.0 Hz), 2.35(1H, d, J=3.8 Hz), 2.44 (1H, d, J=3.1 Hz), 2.75 (1H, d, J=8.7 Hz),3.29-3.34 (1H, m), 3.61 (3H, s), 8.03 (3H, bs). LC-MS (ESI) calcd forC₉H₁₅NO₂ (free amine) 169.11. found 170.3 [M+H⁺] (100%), 339.3 [2M+H⁺](50%).

d) Methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1R,2S,3R,4S)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (0.5 g, 2.43 mmol) was dissolved in methanol (12 mL).Sodium acetate (0.4 g, 4.86 mmol) was added followed by 4 Å powderedmolecular sieves (0.5 g) and 4-fluoro-benzaldehyde (0.302 g, 2.43 mmol).Sodium cyanoborohydride (0.305 g, 4.86 mmol) was added and the mixturewas stirred at 25° C. for 3 h. The mixture was poured into ethyl acetate(300 mL) and shaken with saturated aqueous sodium bicarbonate solution(200 mL). Both layers were passed through a plug of Celite. The organiclayer was further washed with saturated aqueous sodium bicarbonatesolution (100 mL), saturated aqueous brine solution (100 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford thecrude product, methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.675 g, 2.43 mmol, 99%), as a clear oil. LC-MS (ESI) calcd forC₁₆H₂₀FNO₂ 277.15, found 278.2 [M+H⁺].

e)N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(0.6 g, 2.16 mmol) was dissolved in anhydrous N,N-dimethylformamide (20mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.72 g, 2.16 mmol) was addedfollowed by N-methylmorpholine (0.5 mL, 4.54 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.435 g,2.27 mmol) was added and the mixture was stirred at 25° C. for 45 min.Triethylamine (0.91 mL, 6.48 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (300 mL) and washed with 1.0M aqueous hydrochloricacid solution (3×300 mL), saturated aqueous brine solution (100 mL),dried over magnesium sulfate, filtered, and concentrated in vacuo toafford a golden oil. Purification by flash column chromatography (Mercksilica gel 60, 40-63 μm, 0 to 0.75% methanol in dichloromethane)afforded the product as white foam. The foam was dissolved in methanol(10 mL) and the product was precipitated by the addition of 1.0M aqueoushydrochloric acid solution (20 mL) while stirring. The solid wascollected by vacuum filtration and further dried in vacuo to afford thedesired product,N-{3-[(1S,2R,7S,8R)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.592 g, 1.06 mmol, 49%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.15-1.22 (2H, m), 1.39-1.61 (4H, m), 2.49-2.55 (1H, m), 2.62-2.63(1H, m), 3.02 (1H, d, J=9.8 Hz), 3.05 (3H, s), 3.52 (1H, d, J=9.3 Hz),4.41 (1H, d, J=15.5 Hz), 4.95 (1H, d, J=15.5 Hz), 7.14 (2H, t, J=8.7Hz), 7.32 (2H, dd, J₁=8.2 Hz, J₂=5.7 Hz), 7.50 (1H, dd, J₁=8.4 Hz,J₂=2.4 Hz), 7.55-7.57 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₅H₂₅FN₄O₆S₂ 560.12, found 561.3 [M+H⁺]. ee=96% [HPLC-analysis:Chiralpak AS-RH 2.1×150 mm, 5 micron at r.t., Solvent A-Solvent B (seetable for gradient), 0.3 mL/min, 312 nm, t1=4.3 min, t2=6.0 min(major)].

Example 8(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) (rac-di-exo)-3-Amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester

To a stirred solution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acid (1.0g, 6.37 mmol) in anhydrous methanol and benzene (1:1, 20 mL), a 2.0Msolution of (trimethylsilyl)diazomethane in diethyl ether (6.37 mL, 12.7mmol) was added dropwise. The resulting mixture was stirred for 1 h, andconcentrated in vacuo to afford the desired product,(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (1.02 g, 5.85 mmol, 94%), as a colorless oil. ¹H NMR (400MHz, CDCl₃) δ: 1.53 (2H, m), 1.62 (2H, m), 2.68 (1H, d, J=7.6 Hz), 3.27(1H, d, J=7.6 Hz), 3.59 (3H, s), 4.14 (1H, d, J=6.0 Hz), 4.67 (1H, d,J=4.8 Hz).

b)(rac-di-exo)-3-(4-Fluoro-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

4-Fluoro-benzaldehyde (0.62 mL, 5.85 mmol) was added to a solution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (1.02 g, 5.85 mmol) in anhydrous methanol (20 mL) at 25° C.under a nitrogen atmosphere. After stirring for 10 min, glacial aceticacid (0.8 mL) and sodium cyanoborohydride (920 mg, 14.6 mmol) were addedsequentially, and the resulting mixture was stirred at 25° C. for 18 h.The reaction mixture was poured into saturated aqueous sodiumbicarbonate solution and extracted with ethyl acetate. The combinedorganic layers were washed with saturated aqueous brine solution, driedover sodium sulfate, filtered, and concentrated in vacuo. The residuewas dried under high vacuum to afford the desired product,(rac-di-exo)-3-(4-fluoro-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (1.40 g, 5.02 mmol, 86%), as a colorless oil. ¹H NMR(400 MHz, CDCl₃) δ: 1.73 (4H, m), 2.86 (1H, d, J=8.0 Hz), 3.15 (1H, d,J=8.0 Hz), 3.68 (1H, d, J=13.6 Hz), 3.73 (3H, s), 3.84 (1H, d, J=13.6Hz), 4.46 (1H, d, J=5.2 Hz), 4.73 (1H, d, J=4.8 Hz), 6.99 (2H, m), 7.26(2H, m).

c)(rac-di-exo)-3-{(4-Fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

To a stirred solution of(rac-di-exo)-3-(4-fluoro-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (190 mg, 0.68 mmol) in anhydrous N,N-dimethylformamide(4 mL) under a nitrogen atmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 185 mg, 0.55 mmol),N-methylmorpholine (149 μL, 1.36 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (130 mg,0.68 mmol) were added sequentially. After stirring at 25° C. for 3 h,the reaction mixture was poured into 1.0M aqueous hydrochloric acidsolution, and extracted with ethyl acetate. The combined organic layerswere washed with saturated aqueous brine solution, dried over sodiumsulfate, filtered, and concentrated in vacuo. The crude product,(rac-di-exo)-3-{(4-fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester, was used in the next step without furtherpurification. LC-MS (ESI) calcd for C₂₅H₂₇FN₄O₈S₂ 594.13, found 595.2[M+H⁺].

d)(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a stirred solution of the crude(rac-di-exo)-3-{(4-fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester in absolute ethanol (8 mL) under a nitrogenatmosphere, a 21 wt % solution of sodium ethoxide in ethanol (0.81 mL,1.10 mmol) was added. The mixture was stirred at 60° C. for 30 min, andthen cooled to 25° C. 1.0M aqueous hydrochloric acid solution (4 mL, 4mmol) was added slowly to the mixture, upon which a white solidprecipitated. The suspension was stirred for 15 min, filtered through afilter funnel, and washed with water. The solid was collected, driedunder high vacuum to afford analytically pure(rac-di-exo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(165 mg, 0.29 mmol, 53% over two steps), as an off-white solid. ¹H NMR(400 MHz, DMSO-d₆) δ: 1.67 (4H, m), 3.05 (3H, s), 3.32 (1H, m), 3.79(1H, d, J=9.2 Hz), 4.39 (1H, d, J=15.6 Hz), 4.69 (1H, d, J=4.4 Hz), 4.76(1H, d, J=3.6 Hz), 5.04 (1H, d, J=15.6 Hz), 7.16 (2H, m), 7.33 (2H, m),7.49 (1H, dd, J=9.2 Hz), 7.56 (2H, m), 10.17 (1H, s), 13.89 (1H, s).LC-MS (ESI) calcd for C₂₄H₂₃FN₄O₇S₂ 562.10, found 563.4 [M+H⁺].

Example 9(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) (rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acidmethyl ester hydrochloride

(rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acidhydrochloride (1 g, 5.27 mmol) was dissolved in methanol (7 mL). Benzene(10 mL) was added followed by the dropwise addition of a 2.0M solutionof (trimethylsilyl)diazomethane in dichloromethane (5 mL, 10 mmol). Theyellow solution was stirred at 25° C. for 10 min. Additional 2.0Msolution of (trimethylsilyl)diazomethane in dichloromethane (2 mL, 4mmol) was added. The yellow solution was stirred at 25° C. for 10 min.The solution was concentrated in vacuo to afford a yellow oil. The oilwas dissolved in methanol (15 mL) and concentrated in vacuo to affordthe desired product,(rac-di-exo)-3-amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid methylester hydrochloride (1.07 g, 5.25 mmol, 99%), as a yellow oil. LC-MS(ESI) calcd for C₉H₁₃NO₂ (free amine) 167.09, found 168.2 [M+H⁺] (100%),335.4 [2M+H⁺] (25%).

b)(rac-di-exo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

(rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid methylester hydrochloride (1.07 g, 5.25 mmol) was suspended in methanol (23mL). Sodium acetate (0.865 g, 10.54 mmol) was added followed by 4 Åpowdered molecular sieves (1 g) and 4-fluoro-benzaldehyde (0.621 g, 5mmol). Sodium cyanoborohydride (0.662 g, 10.54 mmol) was added and themixture was stirred at 25° C. for 16 h. The mixture was poured into a1:1 mixture of saturated aqueous sodium bicarbonate solution (200 mL)and ethyl acetate (200 mL). The mixture was shaken and the layersseparated. The organic layer was further washed with saturated aqueousbrine solution (50 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(rac-di-exo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (1.044 g, 3.79 mmol, 76%), as a clear oil. LC-MS (ESI)calcd for C₁₆H₁₈FNO₂ 275.13, found 276.2 [M+H⁺].

c)(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(rac-di-exo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (0.083 g, 0.3 mmol) was dissolved in anhydrousN,N-dimethylformamide (2.8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.3 mmol) was addedfollowed by N-methylmorpholine (0.07 mL, 0.63 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.061 g,0.315 mmol) was added and the mixture was stirred at 25° C. for 45 min.Triethylamine (0.126 mL, 0.9 mmol) was added and the mixture was stirredat 50° C. for 16 h. After cooling to 25° C., the solution was dilutedwith a 1.0M aqueous hydrochloric acid solution (8 mL). The resultingprecipitate was collected by vacuum filtration, dissolved in methanoland concentrated in vacuo to afford the crude product as a beige powder.Purification by flash column chromatography (Merck silica gel 60, 40-63μm, 0 to 0.75% methanol in dichloromethane) afforded the product as awhite foam. The foam was triturated with a 1:1 mixture of diethyl etherand hexanes (6 mL) and the resulting solid was collected by vacuumfiltration. The solid was dried in vacuo for 16 h to afford the desiredproduct,(rac-di-exo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(40.3 mg, 0.072 mmol, 24%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.37 (1H, d, J=9.5 Hz), 1.62 (1H, d, J=9.4 Hz), 2.81-2.89 (1H, m),3.05 (3H, s), 3.19-3.40 (4H, m), 4.52 (1H, d, J=15.5 Hz), 5.04 (1H, d,J=14.8 Hz), 6.13 (1H, dd, J₁=5.5 Hz, J₂=3.1 Hz), 6.35 (1H, dd, J₁=5.9Hz, J₂=2.5 Hz), 7.14 (2H, t, J=9.1 Hz), 7.34 (2H, dd, J₁=7.8 Hz, J₂=5.6Hz), 7.49 (1H, dd, J₁=8.8 Hz, J₂=2.0 Hz), 7.54-7.59 (2H, m), 10.16 (1H,s). LC-MS (ESI) calcd for C₂₅H₂₃FN₄O₆S₂ 558.10, found 559.1 [M+H⁺].

Example 10(rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-endo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid ethyl ester

(rac-di-endo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid ethylester hydrochloride (1 g, 4.6 mmol) was suspended in methanol (23 mL).Sodium acetate (0.753 g, 9.18 mmol) was added followed by 4 Å powderedmolecular sieves (1 g) and 4-fluoro-benzaldehyde (0.57 g, 4.59 mmol).Sodium cyanoborohydride (0.577 g, 9.18 mmol) was added and the mixturewas stirred at 25° C. for 16 h. The mixture was poured into a 1:1mixture of saturated aqueous sodium bicarbonate solution (200 mL) andethyl acetate (200 mL). The mixture was shaken and the layers separated.The organic layer was further washed with saturated aqueous brinesolution (50 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(rac-di-endo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (1.18 g, 4.08 mmol, 88%), as a clear oil. LC-MS (ESI)calcd for C₁₇H₂₀FNO₂ 289.15, found 290.2 [M+H⁺].

b)(rac-di-endo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(rac-di-endo)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (0.087 g, 0.3 mmol) was dissolved in anhydrousN,N-dimethylformamide (2.8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.1 g, 0.3 mmol) was addedfollowed by N-methylmorpholine (0.07 mL, 0.63 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.061 g,0.315 mmol) was added and the mixture was stirred at 25° C. for 45 min.Triethylamine (0.126 mL, 0.9 mmol) was added and the mixture was stirredat 50° C. for 16 h. After cooling to 25° C., the solution was dilutedwith a 1.0M aqueous hydrochloric acid solution (8 mL, 8 mmol). Theresulting precipitate was collected by vacuum filtration, dissolved inmethanol and concentrated in vacuo to afford the crude product as abeige powder. Purification by flash column chromatography (Merck silicagel 60, 40-63 μm, 0 to 0.75% methanol in dichloromethane) afforded theproduct as white foam. The foam was triturated with a 1:1 mixture ofdiethyl ether and hexanes (6 mL) and the resulting solid was collectedby vacuum filtration. The solid was dried in vacuo for 16 h to affordthe desired product,(rac-di-endo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(69.3 mg, 0.124 mmol, 41%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.28-1.28 (2H, m), 3.33-3.41 (3H, m), 3.92-4.00 (1H, m), 4.33 (1H, d,J=14.7 Hz), 4.96 (1H, d, J=15.6 Hz), 5.89-5.92 (1H, m), 6.11-6.13 (1H,m), 7.10 (2H, t, J=9.1 Hz), 7.35 (2H, dd, J₁=8.2 Hz, J₂=5.9 Hz), 7.43(1H, dd, J₁=8.4 Hz, J₂=2.4 Hz), 7.48-7.50 (2H, m), 10.10 (1H, s). LC-MS(ESI) calcd for C₂₅H₂₃FN₄O₆S₂ 558.10, found 559.0 [M+H⁺].

Example 11N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(Methoxycarbonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylicacid

exo-4,10-Dioxa-tricyclo[5.2.1.0^(2,6)]decane-3,5-dione (5.10 g, 30.3mmol) was dissolved in a 1:1 mixture of toluene and carbon tetrachloride(600 mL). The mixture was cooled to −55° C. under a nitrogen atmosphere,and then quinine (10.54 g, 32.5 mmol) was added. Methanol (3.59 mL, 90mmol) in a 1:1 mixture of toluene and carbon tetrachloride (30 mL) wasslowly added via an addition funnel. The suspension was stirred at −55°C. for 60 h, and then allowed to warm to 25° C. The mixture wasconcentrated in vacuo and the residue was dissolved in ethyl acetate(400 mL), washed with a 1.0M aqueous hydrochloric acid solution (2×300mL) and saturated aqueous brine solution, dried over magnesium sulfateand filtered. The filtrate was concentrated in vacuo to afford thedesired product,(1S,2R,3S,4R)-3-(methoxycarbonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylicacid (2.46 g, 12.3 mmol, 41%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.49-1.53 (4H, m), 2.99 (2H, s), 3.50 (3H, s), 4.66 (2H, m), 12.15(1H, s).

b) Methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}-7-oxabicyclo[2.2.1]heptane-2-carboxylate

(1S,2R,3S,4R)-3-(Methoxycarbonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylicacid (2.46 g, 12.3 mmol) was dissolved in anhydrous tetrahydrofuran (35mL) and cooled to −10° C. under a nitrogen atmosphere. Triethylamine(5.13 mL, 36.9 mmol) was added followed by the dropwise addition ofethyl chloroformate (2.35 mL, 24.6 mmol) with vigorous stirring.Immediate precipitation was observed. The mixture was stirred at −10° C.for 1 h. Sodium azide (2.40 g, 36.9 mmol) was dissolved in water (17 mL)and added to the reaction mixture at −10° C. The mixture was stirred at−10° C. for 15 min, and then was allowed to warm to 25° C. and stirredfor 2 h. The mixture was poured into water (100 mL) and extracted withethyl acetate. The combined organic layers were washed with saturatedaqueous sodium bicarbonate solution and saturated aqueous brinesolution, dried over magnesium sulfate and filtered. The filtrate wasconcentrated in vacuo to afford the acyl azide intermediate as a clearoil. The oil was dissolved in anhydrous benzene (80 mL) and refluxed for2 h under a nitrogen atmosphere. The solution was allowed to cool to 25°C., and concentrated in vacuo to afford a yellow oil. The oil wasdissolved in dichloromethane (45 mL), triethylamine (3.46 mL, 24.6 mmol)and benzyl alcohol (1.27 mL, 12.3 mmol) were added sequentially. Theresulting mixture was refluxed for 16 h under a nitrogen atmosphere. Themixture was allowed to cool to 25° C., concentrated in vacuo and theresidue was purified by flash column chromatography (Teledyne IscoRediSep column; 0 to 50% ethyl acetate in hexanes) to afford the desiredproduct, methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}-7-oxabicyclo[2.2.1]heptane-2-carboxylate(2.23 g, 7.30 mmol, 59%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:1.51 (2H, m), 1.72 (1H, m), 1.79 (1H, m), 2.97 (1H, d, J=8.4 Hz), 3.56(3H, s), 4.33 (1H, m), 4.37 (d, 1H, J=5.6 Hz), 4.78 (1H, d, J=4.4 Hz),5.10 (2H, m), 5.42 (1H, d, J=10.0 Hz), 7.35 (5H, m). LC-MS (ESI) calcdfor C₁₆H₁₉NO₅ 305.1, found 306.5 [M+H⁺].

c) Methyl (1R,2S,3R,4S)-3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate

To a solution of methyl(1R,2S,3R,4S)-3-{[(benzyloxy)carbonyl]amino}-7-oxabicyclo[2.2.1]heptane-2-carboxylate(2.23 g, 7.30 mmol) in ethyl acetate (60 mL), 5% palladium on carbon(0.5 g, 22% by weight) was added. The flask was degassed and backfilledwith hydrogen gas via balloon. The mixture was stirred at 25° C. for 16h, passed through a plug of Celite and rinsed with ethyl acetate. Thefiltrate was concentrated in vacuo to afford the desired product, methyl(1R,2S,3R,4S)-3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate (1.0 g,5.84 mmol, 80%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ: 1.43 (2H,m), 1.67 (1H, m), 1.76 (1H, m), 2.82 (1H, d, J=7.6 Hz), 3.41 (1H, d,J=7.6 Hz), 3.73 (3H, s), 4.28 (1H, d, J=6.0 Hz), 4.81 (1H, d, J=4.8 Hz).

d) Methyl(1R,2S,3R,4S)-3-[(4-fluoro-3-methylbenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate

To a stirred solution of methyl(1R,2S,3R,4S)-3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate (400 mg,2.34 mmol) in methanol (8 mL) under a nitrogen atmosphere,4-fluoro-3-methyl-benzaldehyde (0.29 mL, 2.34 mmol) was added. Themixture was stirred for 10 min, and then acetic acid (0.4 mL) was addedfollowed by sodium cyanoborohydride (368 mg, 5.85 mmol). The resultingmixture was stirred at 25° C. for 16 h, and then poured into a mixtureof saturated aqueous sodium bicarbonate solution (50 mL) and ethylacetate (100 mL). The layers were separated and the aqueous layer wasextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over magnesium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct, methyl(1R,2S,3R,4S)-3-[(4-fluoro-3-methylbenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate(610 mg, 2.08 mmol, 80%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:1.40 (2H, m), 1.76 (2H, m), 2.27 (3H, d, J=2.0 Hz), 2.86 (1H, d, J=7.6Hz), 3.16 (1H, d, J=8.4 Hz), 3.64 (1H, d, J=13.2 Hz), 3.74 (3H, s), 3.79(1H, d, J=13.6 Hz), 4.46 (1H, d, J=5.2 Hz), 4.73 (1H, d, J=4.8 Hz), 6.92(2H, m), 7.07 (2H, m).

e)N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a stirred solution of methyl(1R,2S,3R,4S)-3-[(4-fluoro-3-methylbenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate(100 mg, 0.34 mmol) and(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 114 mg, 0.34 mmol) inanhydrous N,N-dimethylformamide (4 mL) under a nitrogen atmosphere,N-methylmorpholine (0.075 mL, 0.68 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (72 mg, 0.37mmol) were added sequentially. The mixture was stirred at 25° C. for 2.5h, poured into 1.0M aqueous hydrochloric acid solution, and then wasextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over magnesium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the amideintermediate which was used in the next step without furtherpurification.

The above intermediate was dissolved in ethanol (5 mL), a 21 wt. %solution of sodium ethoxide in ethanol (0.5 mL, 1.36 mmol) was added andthe mixture was stirred at 60° C. for 30 min. The reaction mixture wascooled to 0° C., and then 0.3M aqueous hydrochloric acid solution (10mL) was slowly added. The product precipitated upon stirring. The solidwas collected by filtration, rinsed with water, and further purified byprep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, micron,30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct,N-{3-[(1S,2R,7S,8R)-3-(4-fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(60 mg, 0.10 mmol, 31%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.50-1.71 (4H, m), 2.21 (3H, d, J=2.0 Hz), 3.05 (3H, s),3.40 (1H, m), 3.78 (1H, d, J=9.2 Hz), 4.34 (1H, d, J=15.2 Hz), 4.71 (1H,d, J=4.8 Hz), 4.76 (1H, d, J=3.6 Hz), 5.02 (1H, d, J=15.6 Hz), 7.06-7.21(3H, m), 7.49 (1H, dd, J=8.4, 2.4 Hz), 7.56 (2H, m). LC-MS (ESI) calcdfor C₂₅H₂₅FN₄O₇S₂ 576.1, found 577.5 [M+H⁺]. Anal. calcd forC₂₅H₂₅FN₄O₇S₂: C, 52.07; H, 4.37; N, 9.72; found: C, 51.75; H, 4.63; N,9.77.

Example 12(rac-di-exo)-N-{3-[3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-(4-Fluoro-3-methyl-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

4-Fluoro-3-methyl-benzaldehyde (0.14 mL, 1.10 mmol) was added to asolution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (prepared as described in Example 8a, 200 mg, 1.17 mmol) inanhydrous methanol (10 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 20 min, glacial acetic acid (0.3 mL) and sodiumcyanoborohydride (184 mg, 2.93 mmol) were added sequentially, and theresulting mixture was stirred at 25° C. for 18 h. The reaction mixturewas poured into saturated aqueous sodium bicarbonate solution andextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over sodium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct,(rac-di-exo)-3-(4-fluoro-3-methyl-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (167 mg, 0.57 mmol, 49%), as a yellow oil. ¹H NMR (400MHz, CDCl₃) δ: 1.38-1.48 (2H, m), 1.65-1.82 (2H, m), 2.27 (3H, s), 2.89(1H, d, J=7.6 Hz), 3.26 (1H, d, J=8.0 Hz), 3.67 (1H, d, J=13.2 Hz), 3.75(3H, s), 3.86 (1H, d, J=13.2 Hz), 4.66 (1H, d, J=5.2 Hz), 4.71 (1H, d,J=4.4 Hz), 6.93 (1H, t, J=9.6 Hz), 7.13 (1H, m), 7.19 (1H, m). LC-MS(ESI) calcd for C₁₆H₂₀FNO₃ 293.14, found 294.3 [M+H⁺].

b)(rac-di-exo)-N-{3-[3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a stirred solution of(rac-di-exo)-3-(4-fluoro-3-methyl-benzylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (26 mg, 0.08 mmol) in anhydrous N,N-dimethylformamide(2 mL) under a nitrogen atmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 30 mg, 0.09 mmol),N-methylmorpholine (22 L, 0.2 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19 mg, 0.10mmol) were added sequentially. After stirring at 25° C. for 1.5 h, thereaction mixture was diluted with ethyl acetate, washed with 1.0Maqueous hydrochloric acid solution and saturated aqueous brine solution,dried over sodium sulfate and filtered. The filtrate was concentrated invacuo to afford the crude intermediate. The crude amide intermediate wasdissolved in absolute ethanol (5 mL), and a 21 wt. % solution of sodiumethoxide in ethanol (0.13 mL, 0.35 mmol) was added. The mixture wasstirred at 60° C. for 1 h, and then was allowed to cool to 25° C. A 1.0Maqueous hydrochloric acid solution (4 mL) was added slowly to themixture, and a white solid precipitated upon stirring. The solid wascollected by filtration, rinsed with water and dried in vacuo to affordthe desired product,(rac-di-exo)-N-{3-[3-(4-fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(20 mg, 0.034 mmol, 43%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.50-1.70 (4H, m), 2.21 (3H, s), 3.05 (3H, s), 3.78 (1H, d,J=8.8 Hz), 4.33 (1H, d, J=15.2 Hz), 4.70 (1H, d, J=4.4 Hz), 4.76 (1H, d,J=4.0 Hz), 5.03 (1H, d, J=14.8 Hz), 7.04-7.20 (3H, m), 7.49 (1H, m),7.56 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₇S₂ 576.11,found 577.3 [M+H⁺].

Example 13(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-(3,3-Dimethyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

3,3-Dimethyl-butyraldehyde (120 mg, 1.20 mmol) was added to a solutionof (rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (prepared as described in Example 8a, 200 mg, 1.17 mmol) inanhydrous methanol (10 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 20 min, glacial acetic acid (0.3 mL) and sodiumcyanoborohydride (150 mg, 2.38 mmol) were added sequentially, and theresulting mixture was stirred at 25° C. for 18 h. The reaction mixturewas poured into saturated aqueous sodium bicarbonate solution and wasextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over sodium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct,(rac-di-exo)-3-(3,3-dimethyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (300 mg, 1.17 mmol, 100%), as a yellow oil. ¹H NMR(400 MHz, CDCl₃) δ: 0.91 (9H, s), 1.42 (4H, m), 1.70-1.80 (2H, m), 2.46(1H, m), 2.74 (1H, m), 2.89 (1H, d, J=8.4 Hz), 3.26 (1H, d, J=8.0 Hz),3.71 (3H, s), 4.61 (1H, s), 4.70 (1H, m). LC-MS (ESI) calcd forC₁₄H₂₅NO₃ 255.18, found 256.2 [M+H⁺].

b)(rac-di-exo)-3-(3,3-Dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

To a stirred solution of(rac-di-exo)-3-(3,3-dimethyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (200 mg, 0.78 mmol) in anhydrous N,N-dimethylformamide(4 mL) under a nitrogen atmosphere,(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,287 mg, 0.78 mmol), N-methylmorpholine (0.2 mL, 1.72 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (165 mg,0.86 mmol) were added sequentially. After stirring at 25° C. for 2.5 h,the reaction mixture was diluted with ethyl acetate, washed with 1.0Maqueous hydrochloric acid solution and saturated aqueous brine solution,dried over sodium sulfate and filtered. The filtrate was concentrated invacuo to afford the crude amide intermediate. The above intermediate wasdissolved in absolute ethanol (10 mL), and a 21 wt. % solution of sodiumethoxide in ethanol (1.17 mL, 3.16 mmol) was added. The mixture wasstirred at 60° C. for 1 h, and then was allowed to cool to 25° C. A 1.0Maqueous hydrochloric acid solution (4 mL) was added slowly to themixture, and white solid precipitated upon stirring. The solid wascollected by filtration, rinsed with water and dried in vacuo to affordthe desired product,(rac-di-exo)-3-(3,3-dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(200 mg, 0.35 mmol, 45%), as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 0.93 (9H, s), 1.45 (2H, m), 1.63 (2H, m), 1.71 (2H, m), 1.95 (1H, m),3.30 (1H, m), 3.84 (1H, m), 3.89 (1H, m), 4.74 (2H, bs), 7.35 (1H, d,J=8.4 Hz), 7.98 (1H, dd, J=8.4, 1.6 Hz), 8.07 (1H, d, J=1.6 Hz). LC-MS(ESI) calcd for C₂₂H₂₆₁N₃O₅S 571.06, found 572.3 [M+H⁺].

c)(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

A reaction flask was charged with copper (I) iodide (20 mg, 0.11 mmol),sarcosine (N-methyl glycine) (10 mg, 0.11 mmol), methanesulfonamide (83mg, 0.87 mmol),(rac-di-exo)-3-(3,3-dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(100 mg, 0.17 mmol) and potassium phosphate (111 mg, 0.52 mmol). Theflask was degassed and backfilled with nitrogen, and then anhydrousN,N-dimethylformamide (3 mL) was added. The resulting suspension wasvigorously stirred at 100° C. for 16 h, and then allowed to cool to 25°C. The mixture was passed through a plug of Celite and rinsed with 10%methanol/dichloromethane. The filtrate was concentrated in vacuo, andthe residue was purified by prep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA150×21.2 mm, 5 micron, 30%-95% in 7 min @30 mL/min flow rate, 0.05%trifluoroacetic acid in acetonitrile/0.05% trifluoroacetic acid inwater] to afford the desired product,(rac-di-exo)-N-{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(25 mg, 0.046 mmol, 27%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.94 (9H, s), 1.48 (2H, m), 1.63 (2H, m), 1.70 (2H, m), 2.97(1H, m), 3.30 (1H, m), 3.80-3.90 (2H, m), 4.75 (2H, s), 7.49 (1H, dd,J=8.4, 2.4 Hz), 7.56 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₃H₃₀N₄O₇S₂ 538.16, found 539.4 [M+H⁺].

Example 14N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-ylI}-methanesulfonamide

a) (rac-di-exo)-Methyl 3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate

To a stirred solution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acid (1.0g, 6.37 mmol) in a 1:1 mixture of anhydrous methanol and benzene (20mL), a 2.0M solution of (trimethylsilyl)diazomethane in hexanes (6.37mL, 12.7 mmol) was added dropwise. The resulting mixture was stirred for1 h, and concentrated in vacuo to afford the desired product,(rac-di-exo)-methyl 3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate(1.02 g, 5.96 mmol, 94%), as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ:1.53 (2H, m), 1.62 (2H, m), 2.68 (1H, d, J=7.6 Hz), 3.27 (1H, d, J=7.6Hz), 3.59 (3H, s), 4.14 (1H, d, J=6.0 Hz), 4.67 (1H, d, J=4.8 Hz).

b) (rac-di-exo)-Methyl3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate

4-Fluoro-benzaldehyde (0.62 mL, 5.85 mmol) was added to a solution of(rac-di-exo)-methyl 3-amino-7-oxabicyclo[2.2.1]heptane-2-carboxylate(1.02 g, 5.85 mmol) in anhydrous methanol (20 mL) at 25° C. under anitrogen atmosphere. After stirring for 10 min, glacial acetic acid (0.8mL) and sodium cyanoborohydride (920 mg, 14.6 mmol) were addedsequentially, and the resulting mixture was stirred at 25° C. for 18 h.The reaction mixture was poured into saturated sodium bicarbonatesolution and extracted with ethyl acetate. The combined organic layerswere washed with saturated aqueous brine solution, dried over sodiumsulfate and filtered. The filtrate was concentrated in vacuo, and theresidue was dried under high vacuum to afford the desired product,(rac-di-exo)-methyl3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate (1.40g, 5.01 mmol, 86%), as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ:1.30-1.42 (2H, m), 1.73 (2H, m), 2.86 (1H, d, J=8.0 Hz), 3.15 (1H, d,J=8.0 Hz), 3.68 (1H, d, J=13.6 Hz), 3.73 (3H, s), 3.84 (1H, d, J=13.6Hz), 4.46 (1H, d, J=5.2 Hz), 4.73 (1H, d, J=4.8 Hz), 6.99 (2H, m), 7.26(2H, m).

c) Methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate

To a stirred solution of (rac-di-exo)-methyl3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate (210mg, 0.75 mmol) in ethyl acetate (6 mL), a solution of (S)-(+)-mandelicacid (57.2 mg, 0.38 mmol) in ethyl acetate (3 mL) was added dropwise.The clear solution became cloudy and turned into a suspension uponstirring for 15 min, and stirring was continued for 20 min. The solidwas collected by filtration, rinsed with ethyl acetate and dried invacuo to afford the desired product in a mandelic acid salt form (90 mg,0.21 mmol, 56%) (>96% de, based on ¹H NMR analysis), as a white solid.¹H NMR (400 MHz, CDCl₃) δ: 1.37-1.49 (2H, m), 1.75 (2H, m), 2.88 (1H, d,J=8.0 Hz), 3.78 (1H, d, J=7.6 Hz), 3.65 (1H, d, J=12.8 Hz), 3.72 (3H,s), 3.91 (1H, d, J=13.2 Hz), 4.62 (1H, d, J=5.2 Hz), 4.70 (1H, d, J=4.8Hz), 5.12 (1H, s), 6.94 (2H, m), 7.19 (2H, m), 7.34 (3H, m), 7.46 (2H,m).

The above obtained intermediate (90 mg, 0.21 mmol) was suspended in a1:1 mixture of saturated aqueous sodium bicarbonate solution (5 mL) andethyl acetate (5 mL), and stirred for 30 min at 25° C. The layers wereseparated and the aqueous layer was extracted with ethyl acetate. Thecombined organic layers were washed with saturated aqueous brinesolution, dried over magnesium sulfate and filtered. The filtrate wasconcentrated in vacuo to afford the free amine, methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate(56 mg, 0.21 mmol, 95%), as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:1.25-1.44 (2H, m), 1.64-1.82 (2H, m), 2.86 (1H, d, J=8.0 Hz), 3.15 (1H,d, J=8.0 Hz), 3.67 (1H, d, J=13.6 Hz), 3.73 (3H, s), 3.84 (1H, d, J=13.6Hz), 4.46 (1H, d, J=5.2 Hz), 4.72 (1H, d, J=4.8 Hz), 6.99 (2H, m), 7.26(2H, m).

d)N-{3-[(1S,2R,7S,8R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a stirred solution of methyl(1R,2S,3R,4S)-3-[(4-fluorobenzyl)amino]-7-oxabicyclo[2.2.1]heptane-2-carboxylate(0.056 g, 0.21 mmol) in anhydrous N,N-dimethylformamide (3 mL) under anitrogen atmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.070 g, 0.21 mmol) was addedfollowed by N-methylmorpholine (0.046 mL, 0.42 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (40 mg, 0.21mmol). The mixture was stirred at 25° C. for 1 h, triethylamine (0.88mL, 0.63 mmol) was added and the resulting mixture was stirred at 50° C.for 16 h. The reaction mixture was allowed to cool to 25° C., dilutedwith ethyl acetate, washed with 1.0M aqueous hydrochloric acid solutionand saturated aqueous brine solution, dried over magnesium sulfate andfiltered. The filtrate was concentrated in vacuo and the residue waspurified by prep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5micron, 30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroaceticacid in acetonitrile/0.05% trifluoroacetic acid in water] to afford thedesired product,N-{3-[(1S,2R,7S,8R)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(48 mg, 0.085 mmol, 41%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.51-1.71 (4H, m), 3.05 (3H, s), 3.30 (1H, m), 3.79 (1H, d,J=8.4 Hz), 4.40 (1H, d, J=15.6 Hz), 4.70 (1H, d, J=4.8 Hz), 4.76 (1H, d,J=4.8 Hz), 5.04 (1H, d, J=14.8 Hz), 7.15 (2H, t, J=8.4 Hz), 7.33 (2H,m), 7.49 (1H, dd, J=8.4, 2.4 Hz), 7.55 (2H, m). LC-MS (ESI) calcd forC₂₄H₂₃FN₄O₇S₂ 562.1, found 563.5 [M+H⁺]. Anal. calcd for C₂₄H₂₃FN₄O₇S₂:C, 51.24; H, 4.12; N, 9.96; found: C, 51.10; H, 4.51; N, 9.98. ee>98%[HPLC-analysis: Chiralpak AS-RH 4.6×250 mm, 5 micron, 0.8 mL/min, 310nm].

Example 15N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) Methyl(1S,2R,3S,4R)-3-(4-fluoro-3-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1S,2R,3S,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (prepared as described in Example 6c, 0.32 g, 1.56 mmol)was dissolved in methanol (8 mL). Sodium acetate (0.26 g, 3.12 mmol) wasadded followed by 4 Å powdered molecular sieves (0.32 g) and4-fluoro-3-methyl benzaldehyde (0.19 mL, 1.56 mmol). Sodiumcyanoborohydride (0.24 g, 3.12 mmol) was added and the mixture wasstirred at 25° C. for 16 h. The mixture was poured into a mixture ofsaturated aqueous sodium bicarbonate solution (20 mL) and ethyl acetate(30 mL). After shaking, both layers were passed through a plug ofCelite. The organic layer was further washed with saturated aqueoussodium bicarbonate solution (10 mL), saturated aqueous brine solution(10 mL), dried over magnesium sulfate, filtered, and concentrated invacuo to afford the crude product, methyl(1S,2R,3S,4R)-3-(4-fluoro-3-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate(0.35 g, 1.20 mmol, 77%), as a clear oil. LC-MS (ESI) calcd forC₁₇H₂₂FNO₂ 291.36, found 292.5 [M+H⁺].

b)N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1S,2R,3S,4R)-3-(4-fluoro-3-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate(0.090 g, 0.31 mmol) was dissolved in anhydrous N,N-dimethylformamide (3mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.10 g, 0.31 mmol) was addedfollowed by N-methylmorpholine (0.071 mL, 0.65 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.062 g,0.32 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.13 mL, 0.92 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (50 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×50 mL), saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 50 to 100% ethyl acetate in hexanes) afforded thedesired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.12 g, 0.21 mmol, 68%), as a white powder. ¹H NMR (400 MHz, CDCl₃) δ:1.43-2.85 (9H, m), 2.29 (3H, s), 3.07 (3H, s), 3.45-3.47 (1H, m),5.17-5.21 (2H, m), 6.95-7.05 (3H, m), 7.59-7.66 (3H, m). LC-MS (ESI)calcd for C₂₆H₂₇FN₄O₆S₂ 574.64, found 575.3 [M+H⁺].

Example 16N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) Methyl(1S,2R,3S,4R)-3-(3,4-difluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate

Methyl (1S,2R,3S,4R)-3-aminobicyclo[2.2.1]heptane-2-carboxylatehydrochloride (prepared as described in Example 6c, 0.32 g, 1.58 mmol)was dissolved in methanol (8 mL). Sodium acetate (0.26 g, 3.16 mmol) wasadded followed by 4 Å powdered molecular sieves (0.33 g) and3,4-difluoro benzaldehyde (0.17 mL, 1.58 mmol). Sodium cyanoborohydride(0.22 g, 3.16 mmol) was added and the mixture was stirred at 25° C. for16 h. The mixture was poured into a mixture of saturated aqueous sodiumbicarbonate solution (20 mL) and ethyl acetate (30 mL). After shaking,both layers were passed through a plug of Celite. The organic layer wasfurther washed with saturated aqueous sodium bicarbonate solution (10mL), saturated aqueous brine solution (10 mL), dried over magnesiumsulfate, filtered, and concentrated in vacuo to afford the crudeproduct, methyl(1S,2R,3S,4R)-3-(3,4-difluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate(0.36 g, 1.22 mmol, 78%), as a clear oil. LC-MS (ESI) calcd forC₁₆H₁₉F₂NO₂ 295.32, found 296.3 [M+H⁺].

b)N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methyl(1S,2R,3S,4R)-3-(4-fluoro-3-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylate(0.088 g, 0.30 mmol) was dissolved in anhydrous N,N-dimethylformamide (3mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.10 g, 0.30 mmol) was addedfollowed by N-methylmorpholine (0.069 mL, 0.63 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.060 g,0.32 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.12 mL, 0.90 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (40 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×20 mL), saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 50 to 100% ethyl acetate in hexanes) afforded thedesired product,N-{3-[(1R,2S,7R,8S)-3-(3,4-difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.082 g, 0.14 mmol, 47%), as a white powder. ¹H NMR (400 MHz, CDCl₃) δ:1.42-1.79 (6H, m), 2.51-2.54 (1H, m), 2.84-2.88 (2H, m), 3.07 (3H, s),3.47-3.49 (1H, m), 5.06-5.10 (2H, m), 6.95-7.18 (3H, m), 7.60-7.66 (3H,m). LC-MS (ESI) calcd for C₂₅H₂₄F₂N₄O₆S₂ 578.61, found 579.2 [M+H⁺].

Example 17N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

N-{3-[(1R,2S,7R,8S)-3-(3,4-Difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 16, 114 mg, 0.20 mmol) was dissolvedin N,N-dimethylformamide (7 mL). Potassium carbonate (55 mg, 0.40 mmol)and iodomethane (0.014 mL, 0.22 mmol) were added sequentially. Thereaction was stirred at 25° C. for 18 h. The reaction was quenched viathe addition of 1.0M aqueous hydrochloric acid solution (20 mL). Themixture was extracted with ethyl acetate (3×30 mL). The combined organiclayers were washed with saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 20% ethyl acetate in dichloromethane) afforded thedesired product,N-{3-[(1R,2S,7R,8S)-3-(3,4-difluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide(94 mg, 0.06 mmol, 77%) as a white powder. ¹H NMR (400 MHz, CDCl₃) δ:1.42-1.76 (6H, m), 2.51-2.54 (1H, m), 2.84-2.88 (2H, m), 3.07 (3H, s),3.38 (3H, s), 3.47-3.49 (1H, m), 5.07-5.10 (2H, m), 6.96-7.16 (3H, m),7.72-8.01 (3H, m). LC-MS (ESI) calcd for C₂₆H₂₆F₂N₄O₆S₂ 592.63, found593.4 [M+H⁺].

Example 18N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 15, 92 mg, 0.16 mmol) was dissolved inN,N-dimethylformamide (6 mL). Potassium carbonate (44 mg, 0.32 mmol) andiodomethane (0.011 mL, 0.18 mmol) were added sequentially. The reactionwas stirred at 25° C. for 18 h. The reaction was quenched via theaddition of 1.0M aqueous hydrochloric acid solution (20 mL). The mixturewas extracted with ethyl acetate (3×30 mL). The combined organic layerswere washed with saturated aqueous brine solution (20 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 20% ethyl acetate in dichloromethane) afforded thedesired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-3-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide(91 mg, 0.15 mmol, 96%), as a white powder. ¹H NMR (400 MHz, CDCl₃) δ:1.18-2.85 (9H, m), 2.90 (3H, s), 2.97 (3H, s), 3.38 (3H, s), 3.45-3.47(1H, m), 5.17-5.21 (2H, m), 6.96-7.08 (3H, m), 7.72-8.02 (3H, m). LC-MS(ESI) calcd for C₂₇H₂₉FN₄O₆S₂ 588.67, found 589.2 [M+H⁺].

Example 19(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

N,N-diisopropylethylamine (1.79 mL, 10.3 mmol) and(benzotriazol-1yloxy)-tris(dimethylamino)-phosphoniumhexafluorophosphate(1.52 g, 3.44 mmol) were added sequentially to a solution of(1S,2R,3S,4R)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 6l, 1.0 g, 3.43 mmol)and(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,1.26 g, 3.44 mmol) in N,N-dimethylformamide (25 mL) at 25° C. Theresulting solution was stirred at 25° C. for 19 h, and then wasconcentrated in vacuo. The residue was partitioned between 1.0M aqueoushydrochloric acid solution (150 mL) and ethyl acetate (2×150 mL). Theorganic layers were dried over sodium sulfate, filtered and concentratedin vacuo. The resulting orange oil was dissolved in ethanol (50 mL) at25° C. A 21 wt. % solution of sodium ethoxide in ethanol (3.33 mL, 10.3mmol) was added and the reaction mixture was heated at 60° C. for 3 h.After cooling to 25° C., the reaction mixture was concentrated in vacuoto approximately 5 mL volume and was then partitioned between 1.0Maqueous hydrochloric acid solution (150 mL) and ethyl acetate (2×150mL). The organic layers were dried over sodium sulfate, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (Teledyne Isco RediSep column; 0 to 60% ethyl acetate inhexanes) to afford the desired product,(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(1.0 g, 1.69 mmol, 49%), as a pale yellow foam. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.15-1.21 (3H, m), 1.38-1.41 (1H, m), 1.46-1.61 (3H, m),2.62 (1H, d, J=2.4 Hz), 2.98 (1H, d, J=9.4 Hz), 3.52 (1H, d, J=9.3 Hz),4.40 (1H, d, J=15.7 Hz), 4.95 (1H, d, J=15.6 Hz), 7.14 (2H, d, J=17.9Hz), 7.11-7.16 (2H, m), 7.30-7.34 (3H, m), 7.97 (1H, dd, J₁=2.3 Hz,J₂=8.6 Hz), 8.07 (1H, d, J=1.5 Hz). LC-MS (ESI) calcd for C₂₄H₂₁FIN₃O₄S593.03, found 594.2 [M+H⁺].

Example 20(1R,2S,7R,8S)-5-[7-(1,1-Dioxo-4,5-dihydro-1H-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

To a solution of(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 200 mg, 0.34 mmol) andtributyl-(1,1-dioxo-4,5-dihydro-1H-1λ⁶-thiophen-2-yl)-stannane (preparedas described in US patent application US 2008/0031852, 180 mg, 0.44mmol) in anhydrous N,N-dimethylformamide (7 mL) under a nitrogenatmosphere, Pd(PPh₃)₄ (20 mg, 0.017 mmol) was added. The resultingmixture was stirred at 90° C. for 22 h, and then allowed to cool to 25°C. The reaction mixture was concentrated in vacuo and the residue waspurified by prep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5micron, 30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroaceticacid in acetonitrile/0.05% trifluoroacetic acid in water] to afford thedesired product,(1R,2S,7R,8S)-5-[7-(1,1-dioxo-4,5-dihydro-1H-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(38 mg, 0.065 mmol, 20%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.19 (2H, m), 1.42-1.61 (4H, m), 2.50 (1H, m), 2.65 (1H, m),2.97 (2H, m), 3.05 (1H, m), 3.53 (3H, m), 4.43 (1H, d, J=14.4 Hz), 4.96(1H, d, J=15.6 Hz), 7.15 (2H, m), 7.33 (2H, m), 7.41 (1H, t, J=3.6 Hz),7.66 (1H, d, J=9.2 Hz), 7.96 (1H, dd, J=8.8, 2.4 Hz), 8.10 (1H, d, J=2.4Hz). LC-MS (ESI) calcd for C₂₈H₂₆FN₃O₆S₂ 583.12, found 584.2 [M+H⁺].

Example 21(1R,2S,7R,8S)-5-[7-(1,1-Dioxo-tetrahydro-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

(1R,2S,7R,8S)-5-[7-(1,1-Dioxo-4,5-dihydro-1H-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 20, 30 mg, 0.05 mmol) was dissolved inmethanol (15 mL) and 5% palladium on charcoal (100 mg) was added. Theflask was degassed and backfilled with hydrogen gas via balloon. Themixture was stirred at 25° C. for 16 h. The mixture was passed through aplug of Celite, rinsed with 10% methanol/dichloromethane, and thefiltrate was concentrated in vacuo. The residue was purified byprep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron,30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct,(1R,2S,7R,8S)-5-[7-(1,1-dioxo-tetrahydro-1λ⁶-thiophen-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(26 mg, 0.044 mmol, 86%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.19 (2H, m), 1.40-1.60 (4H, m), 2.12 (1H, m), 2.67 (1H, m), 2.36(1H, m), 2.50 (1H, m), 2.65 (1H, m), 3.03 (1H, m), 3.23 (1H, m), 3.32(1H, m), 3.53 (1H, d, J=10.0 Hz), 4.42 (1H, d, J=15.2 Hz), 4.56 (1H, dd,J=11.6, 6.8 Hz), 4.97 (1H, d, J=15.6 Hz), 7.14 (2H, m), 7.33 (2H, m),7.58 (1H, d, J=8.8 Hz), 7.69 (1H, dd, J=8.8, 2.4 Hz), 7.82 (1H, d, J=2.4Hz); LC-MS (ESI) calcd for C₂₈H₂₈FN₃O₆S₂ 585.14, found 586.3 [M+H⁺].

Example 22(1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(3-Methyl-butylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(1S,2R,3S,4R)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 0.5 g, 2.7 mmol) wasdissolved in methanol (25 mL). Isovaleraldehyde (0.233 g, 2.7 mmol) wasadded followed by acetic acid (1 mL). The solution continued to stir at25° C. for 10 min. Sodium cyanoborohydride (0.424 g, 6.75 mmol) wasadded and the mixture was stirred at 25° C. for 5 h. The mixture waspoured into aqueous half-saturated aqueous sodium bicarbonate solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(1S,2R,3S,4R)-3-(3-methyl-butylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.43 g, 1.7 mmol, 63%), as a light yellow oil, whichwas used in the next step without any further purification. LC-MS (ESI)calcd for C₁₅H₂₇NO₂ 253.2. found 254.1 [M+H⁺].

b)(1S,2R,3S,4R)-3-[[2-(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.216 g, 0.649 mmol) wasdissolved in anhydrous N,N-dimethylformamide (5 mL).(1S,2R,3S,4R)-3-(3-Methyl-butylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.164 g, 0.649 mmol) was added followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.13 g,0.681 mmol). Then N-methylmorpholine (0.138 g, 1.36 mmol) was added intothe above reaction mixture. The mixture was stirred at 25° C. for 4 h.The solution was poured into 1.0M aqueous hydrochloric acid solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over magnisium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(1S,2R,3S,4R)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as a light yellow oil, which was used in the next stepwithout any further purification. LC-MS (ESI) calcd for C₂₅H₃₆N₄O₇S₂568.2, found 569.5 [M+H⁺].

c)(1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

The crude(1S,2R,3S,4R)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester was dissolved in ethanol (7 mL) and a 21 wt. % solutionof sodium ethoxide in ethanol (1.15 mL, 3.25 mmol) was added into theabove solution. The mixture was stirred at 60° C. for 4 h and cooleddown to 25° C. The mixture was poured into 0.5M aqueous hydrochloricacid solution (100 mL). The aqueous layer was extracted with ethylacetate (2×100 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated in vacuo to afford the crude product, whichwas purified by flash column chromatography (Teledyne Isco RediSepcolumn; 100% ethyl acetate) to afford the desired product,(1R,2S,7R,8S)—N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.19 g, 0.364 mmol, 56.1% over two steps), as an off-white solid. ¹HNMR (400 MHz, DMSO-d₆) δ: 0.89-0.93 (6H, m), 1.16-1.23 (2H, m),1.28-1.32 (1H, m), 1.35-1.62 (7H, m), 1.99-1.99 (1H, m), 2.52-2.54 (1H,m), 2.63 (1H, bs), 3.06 (3H, s), 3.62-3.72 (2H, m), 7.50 (1H, dd, J₁=8.9Hz, J₂=2.2 Hz), 7.57-7.59 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₃H₃₀N₄O₆S₂ 522.16, found 523.6 [M+H⁺].

Example 23(rac-di-exo)-N-{3-[3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) Cyclopropylacetaldehyde

To a 2.0M solution of oxalyl chloride in dichloromethane (9.8 mL, 19.6mmol) at −78° C. was added dimethyl sulfoxide dropwise. After stirringfor 15 min at −78° C., a solution of cyclopropylethyl alcohol (1.5 g,17.4 mmol) in dichloromethane (3.5 mL) was added. After stirring for anadditional 1 h, triethylamine (13.8 mL, 98.3 mmol) was added. Thereaction mixture was allowed to warm to 25° C. and diluted with water.The organic layer was separated and the aqueous layer was extracted withdichloromethane (3×50 mL). The combined organic layers were dried overanhydrous magnesium sulfate and concentrated in vacuo at 0° C. to affordthe crude cyclopropylacetaldehyde, which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ 0.19 (2H, dd,J₁=10.3 Hz, J₂=5.2 Hz), 0.62 (2H, dd, J₁=13.2 Hz, J₂=5.3 Hz), 1.03-0.97(1H, m), 2.30 (2H, d, J=5.1 Hz), 9.79 (1H, d, J=1.7 Hz).

b)(rac-di-exo)-3-(2-Cyclopropyl-ethylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

Cyclopropylacetaldehyde (148 mg, 1.75 mmol) was added to a solution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (prepared as described in Example 8a, 300 mg, 1.75 mmol) inanhydrous methanol (10 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 20 min, glacial acetic acid (0.3 mL) and sodiumcyanoborohydride (150 mg, 2.38 mmol) were added sequentially, and theresulting mixture was stirred at 25° C. for 18 h. The reaction mixturewas poured into saturated aqueous sodium bicarbonate solution and wasextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over sodium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct,(rac-di-exo)-3-(2-cyclopropyl-ethylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (220 mg, 0.92 mmol, 53%), as a yellow oil. ¹H NMR (400MHz, CDCl₃) δ: 0.19 (2H, m), 0.52 (2H, m), 0.70 (1H, m), 1.65-2.00 (6H,m), 3.10 (1H, d, J=8.4 Hz), 3.23 (2H, m), 3.82 (3H, s), 3.93 (1H, m),4.90 (1H, d, J=4.8 Hz), 5.17 (1H, d, J=5.2 Hz).

c)(rac-di-exo)-N-{3-[3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a stirred solution of(rac-di-exo)-3-(2-cyclopropyl-ethylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (120 mg, 0.50 mmol) in anhydrous N,N-dimethylformamide(2 mL) under a nitrogen atmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 107 mg, 0.32 mmol),N-methylmorpholine (0.12 mL, 1.09 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (105 mg,0.55 mmol) were added sequentially. After stirring at 25° C. for 2 h,the reaction mixture was diluted with ethyl acetate, washed with 1.0Maqueous hydrochloric acid solution and saturated aqueous brine solution,dried over sodium sulfate and filtered. The filtrate was concentrated invacuo to afford the crude intermediate. The crude amide intermediate wasdissolved in absolute ethanol (5 mL), and a 21 wt. % solution of sodiumethoxide in ethanol (0.8 mL, 2.16 mmol) was added. The mixture wasstirred at 60° C. for 2 h, and then was allowed to cool to 25° C. A 0.5M aqueous hydrochloric acid solution (10 mL) was added, and then themixture was extracted with ethyl acetate, washed with saturated aqueousbrine solution, dried over sodium sulfate and filtered. The filtrate wasconcentrated in vacuo and the residue was purified by prep-HPLC [ColumnLuna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron, 30%-95% in 7 min @30mL/min flow rate, 0.05% trifluoroacetic acid in acetonitrile/0.05%trifluoroacetic acid in water] to afford the desired product,(rac-di-exo)-N-{3-[3-(2-cyclopropyl-ethyl)-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(25 mg, 0.048 mmol, 15%), as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 0.07 (2H, m), 0.41 (2H, m), 0.70 (1H, m), 1.37-1.72 (6H, m), 3.05(3H, s), 3.07 (1H, m), 3.30 (m, 1H), 3.92 (2H, m), 4.74 (2H, m), 7.49(1H, m), 7.55 (2H, m), 10.16 (1H, s). LC-MS (ESI) calcd for C₂₂H₂₆N₄O₇S₂522.12, found 523.4 [M+H⁺].

Example 24(1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

(1R,2S,7R,8S)—N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 22, 90 mg, 0.172 mmol) was dissolvedin anhydrous N,N-dimethylformamide (2 mL). Potassium carbonate (0.04 g,0.344 mmol) was added followed by iodomethane (0.027 g, 0.189 mmol). Themixture was stirred at 25° C. for 5 h. The reaction mixture wasextracted with ethyl acetate (2×100 mL) and water (100 mL). The organiclayer was washed with saturated aqueous brine solution (50 mL), driedover sodium sulfate and filtered. The filtrate was concentrated in vacuoto afford the crude product, which was purified by flash columnchromatography (Teledyne Isco RediSep column; 40% ethyl acetate inhexanes) to afford the desired product,(1R,2S,7R,8S)—N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide(0.052 g, 0.097 mmol, 56.4%), as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.92 (6H, d, J=6.4 Hz), 1.21-1.26 (1H, m), 1.28-1.33 (1H,m), 1.39-1.63 (7H, m), 2.54 (1H, bs), 2.63-2.67 (1H, m), 3.00 (3H, s),3.06-3.17 (1H, m), 3.29 (3H, s), 3.63-3.72 (2H, m), 7.62 (1H, d, J=8.5Hz), 7.70-7.73 (1H, m), 7.85 (1H, s). LC-MS (ESI) calcd for C₂₄H₃₂N₄O₆S₂536.18, found 537.6.6 [M+H⁺].

Example 25(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-(3-Methyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (prepared as described in Example 8a, 0.4 g, 2.34 mmol) wasdissolved in methanol (20 mL). Isovaleraldehyde (0.202 g, 2.34 mmol) wasadded followed by acetic acid (1 mL). The solution continued to stir at25° C. for 10 min. Sodium cyanoborohydride (0.37 g, 5.85 mmol) was addedand the mixture was stirred at 25° C. for 16 h. The mixture was pouredinto a half-saturated aqueous sodium bicarbonate solution (150 mL). Theaqueous layer was extracted with ethyl acetate (2×150 mL). The organiclayer was dried over sodium sulfate, filtered, and concentrated in vacuoto afford the crude product,(rac-di-exo)-3-(3-methyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (0.3 g, 2.34 mmol, 53.1%), as a clear oil. LC-MS (ESI)calcd for C₁₃H₂₃NO₃ 241.17, found 242.4 [M+H⁺].

b)(rac-di-exo)-3-[[2-(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1)⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.211 g, 0.696 mmol) wasdissolved in anhydrous N,N-dimethylformamide (6 mL).(rac-di-exo)-3-(3-Methyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (0.168 g, 0.696 mmol) was added followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.14 g,0.731 mmol). Then N-methylmorpholine (0.148 g, 1.46 mmol) was added intothe above reaction mixture. The mixture was stirred at 25° C. for 5 h.The solution was poured into 1.0M aqueous hydrochloric acid solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester, as a light yellow oil, which was used in the nextstep without any further purification. LC-MS (ESI) calcd forC₂₃H₃₂N₄O₈S₂ 556.17, found 557.4 [M+H⁺].

c)(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester was dissolved in ethanol (7 mL), and a 21 wt. %solution of sodium ethoxide in ethanol (1.3 mL, 3.48 mmol) was addedinto the above solution. The mixture was stirred at 60° C. for 4 h andcooled down to 25° C. The mixture was poured into 0.5M aqueoushydrochloric acid solution (100 mL). The aqueous layer was extractedwith ethyl acetate (2×100 mL). The organic layer was dried over sodiumsulfate, filtered, and concentrated in vacuo to afford the crudeproduct, which was purified by flash column chromatography (TeledyneIsco RediSep column; 100% ethyl acetate) to afford the desired product,(rac-di-exo)-N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide (0.07 g, 0.133 mmol,19.2% over two steps), as a light yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.92 (6H, d, J=5.5 Hz), 1.06-1.31 (2H, m), 1.40-1.72 (6H,m), 3.05 (3H, s), 3.17-3.22 (1H, m), 3.80-3.87 (2H, m), 4.72-4.73 (2H,m), 7.47-7.55 (3H, m), 10.12 (1H, bs). LC-MS (ESI) calcd forC₂₂H₂₈N₄O₇S₂ 524.14, found 525.4 [M+H⁺].

Example 26(1R,2S,7R,8S)-5-[7-(1,1-Dioxo-1λ⁶-isothiazolidin-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

A reaction flask was charged with copper (I) iodide (8 mg, 0.042 mmol),sarcosine (N-methyl glycine) (9 mg, 0.1 mmol), isothiazolidine1,1-dioxide (204 mg, 1.685 mmol),(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 100 mg, 0.168 mmol) and potassiumphosphate (179 mg, 0.842 mmol). The flask was degassed and backfilledwith nitrogen, and then anhydrous N,N-dimethylformamide (3 mL) wasadded. The resulting suspension was vigorously stirred at 100° C. for 17h, and then allowed to cool to 25° C. The mixture was diluted with ethylacetate (30 mL) and washed with 1.0M aqueous hydrochloric acid solution(2×20 mL) and saturated aqueous brine solution (40 mL). The organiclayer was dried over magnesium sulfate, filtered, and concentrated invacuo. Purification by flash column chromatography (Teledyne IscoRediSep column; 1^(st) column: 100% dichloromethane, 2^(nd) column: 5%hexanes in dichloromethane) to afford the desired product. The crudeproduct was triturated with absolute ethanol (3×) and dried in vacuo at60° C. to afford the desired product,(1R,2S,7R,8S)-5-[7-(1,1-dioxo-1λ⁶-isothiazolidin-2-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl]-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(70 mg, 0.119 mmol, 71%), as a solid. ¹H NMR (400 MHz, DMSO-d₆) δ:1.17-1.24 (2H, m), 1.40-1.61 (4H, m), 2.39-2.46 (2H, m), 2.51-2.54 (1H,m), 2.64-2.65 (1H, m), 3.03-3.05 (1H, m), 3.53-3.60 (3H, m), 3.83 (2H,t, J=6.3 Hz), 4.43 (1H, d, J=15.4 Hz), 4.97 (1H, d, J=15.6 Hz), 7.15(2H, t, J=9.0 Hz), 7.32-7.35 (2H, m), 7.51-7.54 (2H, m), 7.62 (1H, d,J=8.5 Hz). LC-MS (ESI) calcd for C₂₇H₂₇FN₄O₆S₂ 586.14, found 587.4[M+H⁺].

Example 27(1R,2S,7R,8S)—N-[3-(6-Hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

A suspension of(1R,2S,3R,4S)-3-ethoxycarbonyl-bicyclo[2.2.1]hept-2-yl-aminium(1′S)-(+)-10-camphorsulfonate (prepared as described in Example 6j, 5.00g, 12.0 mmol) and potassium carbonate (4.16 g, 30.1 mmol) in ethylacetate (80 mL) was stirred at 25° C. for 5 h, then was filtered througha medium frit. The filtrate was concentrated in vacuo to afford thecrude (1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester (2.14 g). A portion of this material (1.00 g, 5.52 mmol) and(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 1.84 g, 5.52 mmol) weredissolved in N,N-dimethylformamide at 25° C. and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.11 g,5.79 mmol) and N-methylmorpholine (1.27 mL, 11.6 mmol) were addedsequentially. The resulting solution was stirred at 25° C. for 22 h, andthen was concentrated in vacuo. The residue was partitioned between 1.0Maqueous hydrochloric acid solution (200 mL) and ethyl acetate (2×150mL). The organic layers were dried over sodium sulfate, filtered andconcentrated in vacuo. The resulting orange oil was dissolved in ethanol(70 mL) at 25° C. A 21 wt. % solution of sodium ethoxide in ethanol(10.7 mL, 33.0 mmol) was added and the reaction mixture was heated at90° C. for 3 d. After cooling to 25° C., the reaction mixture wasconcentrated in vacuo to a volume of approximately 5 mL and was thenpartitioned between 1.0M aqueous hydrochloric acid solution (200 mL) andethyl acetate (2×150 mL). The organic layers were dried over sodiumsulfate, filtered and concentrated in vacuo. The residue was purified byflash column chromatography (Teledyne Isco RediSep column; 0 to 7%methanol in dichloromethane) to afford the desired product,(1R,2S,7R,8S)—N-[3-(6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(0.25 g, 0.552 mmol, 10%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.17-1.22 (2H, m), 1.39-1.58 (3H, m), 2.24 (1H, bs), 2.32 (1H, bs),2.64-2.66 (1H, m), 2.84-2.87 (1H, m), 3.04 (3H, s), 3.54 (1H, bs), 3.63(1H, bs), 7.49 (2H, bs), 7.55 (1H, bs), 8.11 (1H, bs), 9.52 (1H, bs),10.12 (1H, bs). LC-MS (ESI) calcd for C₁₈H₂₀N₄O₆S₂ 452.08, found 453.2[M+H⁺].

Example 28(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a) N-(2-Sulfamoyl-phenyl)-malonamic acid ethyl ester

2-Amino-benzenesulfonamide (5 g, 29 mmol) was dissolved inN,N-dimethylacetamide (25 mL) and diethyl ether (25 mL).Ethyl-3-chloro-3-oxo-propionate (4.6 g, 30.45 mmol) was added into theabove reaction solution. The reaction mixture was stirred at 25° C. for3 h. The product started to precipitate and was collected by vacuumfiltration. The solid was dissolved in ethyl acetate (200 mL) andextracted with water (200 mL). The aqueous layer was back-extracted withethyl acetate (200 mL). The combined organic layers were dried oversodium sulfate, filtered, and concentrated in vacuo to afford the crudeproduct, N-(2-sulfamoyl-phenyl)-malonamic acid ethyl ester, as a whitesolid, which was used in the next step without further purification. ¹HNMR (400 MHz, DMSO-d₆) δ: 1.23 (3H, t, J=7.0 Hz), 3.61 (2H, s), 4.14(2H, quartet, J=7.0 Hz), 7.29-7.33 (1H, m), 7.53 (2H, bs), 7.56-7.60(1H, m), 7.84-7.86 (1H, m), 7.97-7.99 (1H, m), 9.54 (1H, bs). LC-MS(ESI) calcd for C₁₁H₁₄N₂O₅S 286.06, found 287.1 [M+H⁺].

b) (1,1-Dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetic acid

Solid sodium hydroxide (3.48 g, 87 mmol) was dissolved in water to makea saturated solution. The crude N-(2-sulfamoyl-phenyl)-malonamic acidethyl ester was added into the sodium hydroxide solution. The reactionmixture was heated at 110° C. for 2.5 h, and then was cooled down to 25°C. The reaction mixture was acidified by slowly adding a 12.0M aqueoushydrochloric acid solution (9.67 g, 116 mmol) while cooling in anice-water bath. The product precipitated and was collected by vacuumfiltration. The solid was washed with cold water and dried under highvacuum to afford the crude product,(1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetic acid (5g, 20.8 mmol, 71.7% over two steps), as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 3.58 (2H, s), 7.31 (1H, d, J=8.0 Hz), 7.44 (1H, dd, J₁=7.8Hz, J₂=7.8 Hz), 7.67 (1H, dd, J₁=7.8 Hz, J₂=7.8 Hz), 7.79 (1H, d, J=7.9Hz), 12.18 (1H, bs), 13.03 (1H, bs). LC-MS (ESI) calcd for C₉H₈N₂O₄S240.02, found 241.1 [M+H⁺].

c)(1S,2R,3S,4R)-3-[[2-(1,1-Dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(4-fluoro-benzyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(1,1-Dioxo-1,4-dihydro-1□6-benzo[1,2,4]thiadiazin-3-yl)-acetic acid (0.2g, 0.833 mmol) was dissolved in anhydrous N,N-dimethylformamide (8 mL).(1S,2R,3S,4R)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 6l, 0.244 g, 0.833mmol) was added followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.168 g,0.875 mmol). Then N-methylmorpholine (0.177 g, 1.75 mmol) was added intothe above reaction mixture. The mixture was stirred at 25° C. for 16 h.The solution was poured into 1.0M aqueous hydrochloric acid solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(1S,2R,3S,4R)-3-[[2-(1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(4-fluoro-benzyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as an orange oil, which was used in the next stepwithout any further purification. LC-MS (ESI) calcd for C₂₆H₂₈FN₃O₅S513.58, found 514.4 [M+H⁺].

d)(1R,2S,7R,8S)-5-(1,1-Dioxo-1,2-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

The crude(1S,2R,3S,4R)-3-[[2-(1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(4-fluoro-benzyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester was dissolved in ethanol (8 mL), and a 21 wt. %solution of sodium ethoxide in ethanol (1.6 mL, 4.2 mmol) was added intothe above solution. The mixture was stirred at 60° C. for 4 h andallowed to cool down to 25° C. The mixture was poured into 0.5M aqueoushydrochloric acid solution (100 mL). The product started to precipitateand was collected by vacuum filtration. The precipitate was purified byflash column chromatography (Teledyne Isco RediSep column; 100% ethylacetate) to afford the desired product,(1R,2S,7R,8S)-5-(1,1-dioxo-1,2-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.242 g, 0.517 mmol, 62.1% over two steps), as a white solid. ¹H NMR(400 MHz, DMSO-d₆) δ: 1.16-1.22 (2H, m), 1.40-1.60 (4H, m), 2.51 (1H,bs), 2.64 (1H, d, J=2.1 Hz), 3.03 (1H, d, J=8.0 Hz), 3.54 (1H, d, J=9.3Hz), 4.42 (1H, d, J=15.6 Hz), 4.97 (1H, d, J=15.7 Hz), 7.15 (2H, t,J=8.8 Hz), 7.33 (2H, dd, J₁=8.0 Hz, J₂=5.9 Hz), 7.45-7.53 (2H, m),7.67-7.71 (1H, m), 7.85 (1H, d, J=7.9 Hz). LC-MS (ESI) calcd forC₂₄H₂₂FN₃O₄S 467.13, found 468.2 [M+H⁺]. Anal. calcd for C₂₄H₂₂FN₃O₄S:C, 61.66; H, 4.74; N, 8.99; found C, 61.96; H, 4.88; N, 8.99.

Example 29N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

a)(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid

A solution of (7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid ethyl ester (prepared as described in US patent application US2008/0031852, 600 mg, 1.666 mmol) in methanol (30 mL) was cooled to 0°C. in an ice-water bath and treated with 2.0M aqueous lithium hydroxidesolution (4.17 mL, 8.332 mmol). The reaction mixture was allowed to warmto 25° C. and stirred for 1 h. The methanol was removed in vacuo and thereaction mixture was poured into 0.5M aqueous hydrochloric acid solution(20 mL) on ice, extracted with ethyl acetate (3×50 mL), dried oversodium sulfate, filtered, and concentrated in vacuo to afford an orangesolid. The crude solid was triturated with diethyl ether to afford thedesired product,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid (409 mg, 1.232 mmol, 73.9%), as a yellow solid. LC-MS calcd forC₁₁H₁₂N₂O₆S₂ 332.4, found 333.0 [M+H⁺].

b)N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

Methyl(1S,2R,3S,4R)-3-[(4-fluorobenzyl)amino]bicyclo[2.2.1]heptane-2-carboxylate(prepared as described in Example 6d, 0.20 g, 0.69 mmol) was dissolvedin anhydrous N,N-dimethylformamide (7 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid (0.23 g, 0.69 mmol) was added followed by N-methylmorpholine (0.17mL, 1.52 mmol). The mixture was stirred until everything dissolved,approximately 5 min. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (0.15 g, 0.76 mmol) was added and the mixture was stirredat 25° C. for 16 h. The reaction was quenched via addition of saturatedaqueous sodium bicarbonate solution (20 mL). The mixture was extractedwith ethyl acetate (3×30 mL). The organic layers were combined andwashed with saturated aqueous brine solution (20 mL). The resultingsolution was dried over magnesium sulfate, filtered, and concentrated invacuo to afford a golden oil. The oil was dissolved in ethanol (10 mL).A 21 wt. % solution of sodium ethoxide in ethanol (0.65 mL, 1.74 mmol)was added. The reaction was stirred at 60° C. for 16 h. The reaction wasquenched via the addition of 1.0M aqueous hydrochloric acid solution (10mL). The mixture was extracted with ethyl acetate (3×20 mL). The organiclayer was further washed with saturated aqueous sodium bicarbonatesolution (2×20 mL), saturated aqueous brine solution (20 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 22 to 75% ethyl acetate in hexanes) afforded the desiredproduct,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide(0.020 g, 0.04 mmol, 5.3%), as a yellow powder. ¹H NMR (400 MHz, CDCl₃)δ: 1.21-1.64 (6H, m), 2.52-2.71 (3H, m), 3.07 (3H, s), 3.39-3.52 (1H,m), 5.15-5.28 (2H, m), 6.60 (1H, s), 7.02-7.06 (2H, m), 7.22-7.26 (2H,m), 7.54-7.66 (3H, m). LC-MS (ESI) calcd for C₂₆H₂₆FN₃O₆S₂ 559.63, found560.5 [M+H⁺].

Example 30(1R,2S,7R,8S)-5-(7-Amino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a)(1R,2S,7R,8S)-5-(7-Azido-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 0.513 g, 0.864 mmol), sodium azide(1.12 g, 17.2 mmol), sodium ascorbate (0.086 g, 0.43 mmol), copper (I)iodide (0.16 g, 0.84 mmol), andtrans-N,N′-dimethylcyclohexane-1,2-diamine (0.20 mL, 1.27 mmol) weredissolved in a 5:1 mixture of dimethyl sulfoxide and water (10 mL) at25° C. The reaction flask was degassed and backfilled with nitrogen(5×). After stirring at 25° C. for 14 h, the reaction mixture waspartitioned between water (150 mL) and ethyl acetate (2×150 mL). Theorganic layer was dried over sodium sulfate, filtered and concentratedin vacuo. The residue was purified by flash column chromatography(Teledyne Isco RediSep column; 0 to 60% ethyl acetate in hexanes) toafford the desired product,(1R,2S,7R,8S)-5-(7-azido-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.348 g, 0.684 mmol, 79%), as a dark brown foam, which was used in thenext step without any further purification.

b)(1R,2S,7R,8S)-5-(7-Amino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

(1R,2S,7R,8S)-5-(7-Azido-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.348 g, 0.684 mmol) was dissolved in a 1:1 mixture if methanol andethyl acetate (15 mL) at 25° C. Palladium on carbon (0.40 g, 5%, “wet”)was added, resulting in a black suspension. The reaction was maintainedunder a hydrogen atmosphere (balloon) at 25° C. for 6 h, and then wasfiltered through Celite. The Celite was washed with ethyl acetate (2×30mL) and the filtrate was concentrated in vacuo. The residue was purifiedby flash column chromatography (Teledyne Isco RediSep column; 50 to 100%ethyl acetate in hexanes) to afford the desired product,(1R,2S,7R,8S)-5-(7-amino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.159 g, 0.330 mmol, 48%), as a pale yellow solid. ¹H NMR (400 MHz,CDCl₃) δ: 1.08-1.19 (3H, m), 1.40-1.57 (3H, m), 2.99 (1H, d, J=7.2 Hz),3.31 (3H, s), 3.36-3.37 (1H, m), 3.50 (1H, d, J=7.8 Hz), 4.39 (1H, d,J=14.6 Hz), 4.93 (1H, d, J=14.5 Hz), 6.86-6.91 (3H, m), 7.13-7.15 (2H,m), 7.21 (1H, d, J=8.8 Hz), 7.30 (2H, bs), 13.79 (1H, s). LC-MS (ESI)calcd for C₂₄H₂₃FN₄O₄S 482.14, found 483.4 [M+H⁺].

Example 31(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) (rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acidmethyl ester hydrochloride

(rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acidhydrochloride (1.0 g, 5.27 mmol) was dissolved in methanol (7 mL).Benzene (10 mL) was added followed by the dropwise addition of a 2.0Msolution of (trimethylsilyl)diazomethane in diethyl ether (5 mL, 10mmol). After stirring for 10 min, additional 2.0M solution of(trimethylsilyl)diazomethane in diethyl ether (3 mL, 6 mmol) was addedand the reaction mixture was stirred for an additional 20 min afterwhich time the solution remained yellow in color. The solvents wereremoved in vacuo, the residue was taken up in methanol (15 mL) and thesolvent was removed in vacuo to afford the crude product,(rac-di-exo)-3-amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid methylester hydrochloride (0.98 g, 4.83 mmol, 91.6%), as a yellow oil, whichwas used in the next step without any further purification.

b)(rac-di-exo)-3-(3-Methyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

(rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid methylester hydrochloride (0.955 g, 4.703 mmol) was suspended in methanol (10mL). Sodium acetate (0.791 g, 9.652 mmol) was added followed by 4 Åpowdered molecular sieves (1.0 g) and 3-methyl-butyraldehyde (0.385 g,4.468 mmol). Sodium cyanoborohydride (0.593 g, 9.406 mmol) was added andthe mixture was stirred at 25° C. for 18 h. The mixture was poured intoa saturated aqueous sodium bicarbonate solution (100 mL) and extractedwith ethyl acetate (100 mL). The layers were separated and the organiclayer was washed with saturated aqueous brine solution, dried oversodium sulfate, filtered, and concentrated in vacuo. Furtherpurification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 10% methanol in dichloromethane) afforded the desiredproduct,(rac-di-exo)-3-(3-methyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (0.774 g, 3.14 mmol, 66.7%), as a yellow oil. ¹H NMR(400 MHz, CDCl₃) δ: 0.89 (3H, d, J=1.6 Hz), 0.90 (3H, d, J=2.1 Hz),1.27-1.39 (2H, m), 1.54-1.64 (2H, m), 2.07 (1H, d, J=9.4 Hz), 2.42-2.56(2H, m), 2.66-2.73 (1H, m), 2.84 (1H, bs), 2.92 (1H, bs), 2.98 (1H, d,J=7.7 Hz), 3.69 (3H, s), 6.14 (2H, s). LC-MS calcd for C₁₄H₂₃NO₂ 237.17,found 238.2 [M+H⁺].

c)(rac-di-exo)-3-[[2-(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a solution of(rac-di-exo)-3-(3-methyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (208.7 mg, 0.880 mmol) in N,N-dimethylformamide (3 mL)was added(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 351.7 mg, 1.056 mmol) and themixture was vortexed until all material had completely dissolved. A 1.0Msolution of N,N-dicyclohexylcarbodiimide in dichloromethane (1.144 mL,1.144 mmol) was added to the above solution and the mixture was stirredat 25° C. for 18 h under a nitrogen atmosphere. The precipitatedN,N-dicyclohexylurea was removed by filtration over Celite, the filtercake was washed with dichloromethane (3×5 mL), and the solvent wasremoved in vacuo. The residue was dissolved in a 1:1 mixture of ethylacetate and hexanes (20 mL), washed with saturated aqueous brinesolution, saturated aqueous sodium bicarbonate solution and water. Thelayers were separated and the aqueous layers were back-extracted with a1:1 mixture of ethyl acetate and hexanes (20 mL). The combined organiclayers were dried over sodium sulfate, filtered, and concentrated invacuo to afford the crude product,(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, as a yellow oil, which was used in the next stepwithout any further purification. LC-MS calcd for C₂₄H₃₂N₄O₇S₂ 552.17,found 553.3 [M+H⁺].

d)(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester was dissolved in ethanol (20 mL). To this solution wasadded a 21 wt. % solution of sodium ethoxide in ethanol (1.141 g, 3.52mmol) and the reaction mixture was stirred at 25° C. for 2 h. Uponaddition of a 1.0M aqueous hydrochloric acid solution the productprecipitated and was collected by vacuum filtration. Furtherpurification by trituration with dichloromethane and ethyl acetate gavea tan solid, which was further washed with ethyl acetate, and then driedin vacuo, to afford the desired product,(rac-di-exo)-N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(116.6 mg, 0.224 mmol, 25.5%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.93 (6H, d, J=5.3 Hz), 1.35-1.70 (5H, m), 2.85 (1H, d,J=8.7 Hz), 3.06 (3H, s), 3.13-3.31 (5H, m), 3.49 (1H, d, J=8.6 Hz),3.75-3.83 (1H, m), 6.23 (1H, s), 6.39 (1H, s), 7.50-7.58 (3H, m), 10.18(1H, s). LC-MS calcd for C₂₃H₂₈N₄O₆S₂ 520.15, found 521.4 [M+H⁺].

Example 32(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-(3,3-Dimethyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

(rac-di-exo)-3-Amino-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid methylester hydrochloride (0.984 g, 4.846 mmol) was suspended in methanol (10mL). Sodium acetate (0.795 g, 9.692 mmol) was added followed by 4 Åpowdered molecular sieves (1.0 g) and 3,3-dimethylbutyraldehyde (0.461g, 4.604 mmol). Sodium cyanoborohydride (0.610 g, 9.692 mmol) was addedand the mixture was stirred at 25° C. for 18 h. The mixture was pouredinto a saturated aqueous sodium bicarbonate solution (100 mL) andextracted with ethyl acetate (100 mL). The layers were separated and theorganic layer was washed with saturated aqueous brine solution, driedover sodium sulfate, filtered, and concentrated in vacuo. Furtherpurification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 10% methanol in dichloromethane) afforded the desiredproduct,(rac-di-exo)-3-(3,3-dimethyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (0.773 g, 3.08 mmol, 63.5%), as a yellow oil. ¹H NMR(400 MHz, CDCl₃) δ: 0.91 (9H, s), 1.31-1.40 (2H, m), 1.55 (1H, d, J=9.5Hz), 2.08 (1H, d, J=8.8 Hz), 2.43-2.49 (1H, m), 2.55 (1H, d, J=7.8 Hz),2.64-2.74 (1H, m), 2.85 (1H, bs), 2.93 (1H, bs), 2.99 (1H, d, J=7.7 Hz),3.69 (3H, s), 6.15 (2H, s). LC-MS calcd for C₁₅H₂₅NO₂ 251.19, found252.2 [M+H⁺].

b)(rac-di-exo)-3-{(3,3-Dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a solution of(rac-di-exo)-3-(3,3-dimethyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (233.3 mg, 0.929 mmol) in N,N-dimethylformamide (3 mL)was added(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 371.2 mg, 1.115 mmol) and themixture was vortexed until all material had completely dissolved. A 1.0Msolution of N,N-dicyclohexylcarbodiimide in dichloromethane (1.208 mL,1.208 mmol) was added to the above solution and the mixture was stirredat 25° C. for 18 h under a nitrogen atmosphere. The precipitatedN,N-dicyclohexylurea was removed by filtration over Celite, the filtercake was washed with dichloromethane (3×5 mL), and the solvent wasremoved in vacuo. The residue was dissolved in a 1:1 mixture of ethylacetate and hexanes (20 mL), washed with saturated aqueous brinesolution, saturated aqueous sodium bicarbonate solution and water. Thelayers were separated and the aqueous layers were back-extracted with a1:1 mixture of ethyl acetate and hexanes (20 mL). The combined organiclayers were dried over sodium sulfate, filtered, and concentrated invacuo to afford the crude product,(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, as a yellow oil, which was used in the next stepwithout any further purification. LC-MS calcd for C₂₅H₃₄N₄O₇S₂ 566.19,found 567.4 [M+H⁺].

c)(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (710.8 mg, 0.929 mmol) was dissolved in ethanol. Tothis solution was added a 21 wt. % solution of sodium ethoxide inethanol (1.204 g, 3.716 mmol) and the reaction mixture was stirred at25° C. for 2 h. Upon addition of a 1.0M aqueous hydrochloric acidsolution the product precipitated and was collected by vacuumfiltration. Further purification by trituration with dichloromethane andethyl acetate gave a tan solid, which was further washed with ethylacetate, and then dried in vacuo, to afford the desired product,(rac-di-exo)-N-{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(210.7 mg, 0.394 mmol, 42.5% over two steps), as an off-white solid. ¹HNMR (400 MHz, DMSO-d₆) δ: 0.96 (9H, s), 1.39-1.56 (4H, m), 2.85 (1H, d,J=9.3 Hz), 3.06 (3H, s), 3.12-3.30 (5H, m), 3.49 (1H, d, J=9.1 Hz),3.74-3.81 (1H, m), 6.23 (1H, s), 6.40 (1H, s), 7.50-7.60 (3H, m), 10.18(1H, s). LC-MS calcd for C₂₄H₃₀N₄O₆S₂ 534.16, found 535.4 [M+H⁺].

Example 33(rac-di-exo)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a)(rac-di-exo)-3-[[2-(7-Iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

(7-Iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,0.2 g, 0.546 mmol) was dissolved in anhydrous N,N-dimethylformamide (5mL).(rac-di-exo)-3-(3-Methyl-butylamino)-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (prepared as described in Example 25a, 0.132 g, 0.546mmol) was added followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.11 g,0.573 mmol). Then N-methylmorpholine (0.116 g, 1.15 mmol) was added intothe above reaction mixture. The mixture was stirred at 25° C. for 16 h.The solution was poured into 1.0M aqueous hydrochloric acid solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(rac-di-exo)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester, as a light yellow oil, which was used in the nextstep without any further purification. LC-MS (ESI) calcd forC₂₂H₂₈₁N₃O₆S 589.07, found 590.5 [M+H⁺].

b)(rac-di-exo)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

The crude(rac-di-exo)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester was dissolved in ethanol (5 mL), and a 21 wt. %solution of sodium ethoxide in ethanol (1 mL, 2.73 mmol) was added intothe above solution. The mixture was stirred at 60° C. for 6 h andallowed to cool to 25° C. The mixture was poured into 0.5M aqueoushydrochloric acid solution (100 mL). The product started to precipitateand was collected by vacuum filtration. The precipitate was purified byflash column chromatography (Teledyne Isco RediSep column; 100% ethylacetate) to afford the desired product,(rac-di-exo)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.066 g, 0.118 mmol, 21.7% over two steps), as a white solid. ¹H NMR(400 MHz, DMSO-d₆) δ: 0.92 (6H, d, J=6.2 Hz), 1.41-1.73 (7H, m),2.52-2.52 (1H, m), 2.94-3.07 (1H, m), 3.81-3.92 (2H, m), 4.75 (2H, d,J=2.2 Hz), 7.37 (1H, d, J=8.5 Hz), 8.00 (1H, dd, J₁=8.6 Hz, J₂=2.2 Hz),8.08 (1H, s). LC-MS (ESI) calcd for C₂₁H₂₄₁N₃O₅S 557.05, found 558.2[M+H⁺]. Anal. calcd for C₂₁H₂₄₁N₃O₅S.0.5 H₂O: C, 44.53; H, 4.45; N,7.45; found C, 44.69; H, 4.15; N, 7.45.

Example 34(1R,2S,7R,8S)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a)(1S,2R,3S,4R)-3-[[2-(7-Iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(7-Iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,0.2 g, 0.546 mmol) was dissolved in anhydrous N,N-dimethylformamide (5mL).(1S,2R,3S,4R)-3-(3-Methyl-butylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 22a, 0.138 g, 0.546mmol) was added followed by1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.11 g,0.573 mmol). Then N-methylmorpholine (0.116 g, 1.15 mmol) was added intothe above reaction mixture. The mixture was stirred at 25° C. for 16 h.The solution was poured into 1.0M aqueous hydrochloric acid solution(100 mL). The aqueous layer was extracted with ethyl acetate (2×100 mL).The organic layer was dried over sodium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(1S,2R,3S,4R)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as a light yellow oil, which was used in the next stepwithout any further purification. LC-MS (ESI) calcd for C₂₄H₃₂₁N₃O₅S601.11, found 602.6 [M+H⁺].

b)(1R,2S,7R,8S)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

The crude(1S,2R,3S,4R)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.546 mmol) was dissolved in ethanol (5 mL), and a 21wt. % solution of sodium ethoxide in ethanol (1 mL, 2.73 mmol) was addedinto the above solution. The mixture was stirred at 60° C. for 6 h andallowed to cool to 25° C. The mixture was poured into 0.5M aqueoushydrochloric acid solution (100 mL). The product started to precipitateand was collected by vacuum filtration. The precipitate was purified byflash column chromatography (Teledyne Isco RediSep column; 40% ethylacetate in hexanes) to afford the desired product,(1R,2S,7R,8S)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.122 g, 0.2 mmol, 40.3% over two steps), as a light yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ: 0.92 (6H, d, J=5.4 Hz), 1.16-1.63 (9H, m),2.99-3.12 (2H, m), 3.62-3.71 (2H, m), 7.37 (1H, d, J=8.3 Hz), 7.99 (1H,dd, J₁=8.6 Hz, J₂=1.6 Hz), 8.08 (1H, s). LC-MS (ESI) calcd forC₂₂H₂₆₁N₃O₄S 555.07, found 556.3 [M+H⁺].

Example 35 Cyclopropane sulfonic acid{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 81 mg, 0.14 mmol), cyclopropanesulfonic acid amide (132 mg, 1.09 mmol), sarcosine (N-methyl glycine)(18 mg, 0.20 mmol), copper (I) iodide (26 mg, 0.14 mmol), and potassiumphosphate (173 mg, 0.82 mmol) were combined and dissolved inN,N-dimethylformamide (4 mL). The flask was degassed and backfilled withnitrogen (3×). The reaction was stirred at 100° C. for 4 h. The mixturewas allowed to cool to 25° C., diluted with ethyl acetate (20 mL), andextracted with saturated aqueous sodium bicarbonate solution (2×20 mL).The combined organic layers were dried over magnesium sulfate, filteredand concentrated in vacuo. Purification by flash column chromatography(Teledyne Isco RediSep column; 0 to 8% methanol in dichloromethane)afforded the desired product, cyclopropane sulfonic acid{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(76 mg, 0.13 mmol, 96%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆) δ:0.85-1.68 (8H, m), 2.50-2.62 (2H, m), 2.83-2.97 (3H, m), 3.45-3.47 (1H,m), 4.56 (1H, bs), 5.17-5.20 (1H, m), 7.03-7.07 (2H, m), 7.20-7.23 (2H,m), 7.63-7.69 (2H, m), 8.01 (1H, s). LC-MS (ESI) calcd for C₂₇H₂₇FN₄O₆S₂586.14, found 587.4 [M+H⁺].

Example 36(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-{(3,3-Dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a stirred solution of(rac-di-exo)-3-(3,3-dimethyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 32a, 103 mg, 0.410mmol) in anhydrous N,N-dimethylformamide (3 mL) under a nitrogenatmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid (prepared as described in Example 29a, 136 mg, 0.410 mmol),N-methylmorpholine (87.1 mg, 0.861 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (82.3 mg,0.431 mmol) were added sequentially. After stirring at 25° C. for 90min, 1.0M aqueous hydrochloric acid solution (10 mL) and saturatedaqueous brine solution were added to the reaction mixture. The mixturewas extracted with ethyl acetate (3×10 mL) and the combined organiclayers were dried over sodium sulfate, filtered, and concentrated invacuo. The crude product,(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, was directly used in the next step without furtherpurification. LC-MS calcd for C₂₆H₃₅N₃O₇S₂ 565.19, found 566.5 [M+H⁺].

b)(rac-di-exo)-N-{3-[3-(3,3-Dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester was dissolved in ethanol (10 mL). To this solution wasadded a 21 wt. % solution of sodium ethoxide in ethanol (0.399 g, 1.23mmol) and the reaction mixture was stirred at 25° C. for 2.5 h. A 1.0Maqueous hydrochloric acid solution (10 mL) was added and, after stirringfor another 30 min, additional 1.0M aqueous hydrochloric acid solution(5 mL) was added upon which the product precipitated. The solid wascollected by vacuum filtration, washed with 1.0M aqueous hydrochloricacid solution (5 mL), and then dried in vacuo, to afford the desiredproduct,(rac-di-exo)-N-{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide(59 mg, 0.111 mmol, 27.1% over two steps), as a yellow-tan solid. ¹H NMR(400 MHz, CDCl₃) δ: 1.00-1.01 (9H, m), 1.26-1.29 (2H, m), 1.51-1.70 (4H,m), 2.57-2.63 (1H, m), 3.07 (3H, s), 3.10-3.17 (2H, m), 3.24-3.26 (1H,m), 3.35-3.39 (1H, m), 3.39-3.99 (1H, m), 4.90 (0.7H, d, J=16.7 Hz),5.20 (0.3H, d, J=16.2 Hz), 5.27 (0.7H, d, J=16.9 Hz), 5.73 (0.3H, d,J=16.1 Hz), 6.17-6.22 (1H, m), 6.36-6.39 (1H, m), 7.17 (1H, s),7.22-7.25 (2H, m), 7.56-7.61 (1H, m), 7.67-7.69 (1H, m). LC-MS calcd forC₂₅H₃₁N₃O₆S₂ 533.17, found 534.4 [M+H⁺].

Example 37(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-{(4-Fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a stirred solution of(rac-di-exo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 9b, 112.7 mg, 0.410mmol) in anhydrous N,N-dimethylformamide (3 mL) under a nitrogenatmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid (prepared as described in Example 29a, 136 mg, 0.410 mmol),N-methylmorpholine (87.1 mg, 0.861 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (82.3 mg,0.431 mmol) were added sequentially. After stirring at 25° C. for 90min, 1.0M aqueous hydrochloric acid solution (10 mL) and saturatedaqueous brine solution (10 mL) were added to the reaction mixture. Themixture was extracted with ethyl acetate (3×10 mL) and the combinedorganic layers were dried over sodium sulfate, filtered, andconcentrated in vacuo. The crude product,(rac-di-exo)-3-{(4-fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, was used in the next step without any furtherpurification. LC-MS calcd for C₂₇H₂₈FN₃O₇S₂ 589.14, found 590.5 [M+H⁺].

c)(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-{(4-fluoro-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester was dissolved in ethanol. To this solution was added a21 wt. % solution of sodium ethoxide in ethanol (0.399 g, 1.23 mmol) andthe reaction mixture was stirred at 25° C. for 2.5 h. A 1.0M aqueoushydrochloric acid solution (10 mL) was added and, after stirring foranother 30 min, additional 1.0M aqueous hydrochloric acid solution (5mL) was added upon which the product precipitated. The solid wascollected by vacuum filtration, washed with 1.0M aqueous hydrochloricacid solution (5 mL), and then dried in vacuo, to afford the desiredproduct,(rac-di-exo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide(44 mg, 0.079 mmol, 19.3% over two steps), as a tan solid. ¹H NMR (400MHz, CDCl₃) δ: 0.83-0.93 (1H, m), 1.26-1.29 (2H, m), 1.51 (1H, d, J=8.4Hz), 1.69 (1H, d, J=9.5 Hz), 2.59 (1H, d, J=8.3 Hz), 3.05 (3H, s), 3.09(1H, s), 3.26 (1H, s), 3.35 (1H, d, J=7.7 Hz), 4.36 (1H, d, J=15.1 Hz),5.24 (1H, d, J=15.6 Hz), 6.12 (1H, s), 6.33 (1H, s), 7.00-7.04 (2H, m),7.19-7.26 (3H, m), 7.56 (1H, d, J=6.8 Hz), 7.68 (1H, s). LC-MS calcd forC₂₆H₂₄FN₃O₆S₂ 557.11, found 558.1 [M+H⁺].

Example 38(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

a)(rac-di-exo)-3-[[2-(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a stirred solution of(rac-di-exo)-3-(3-methyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 31b, 97.2 mg, 0.410mmol) in anhydrous N,N-dimethylformamide (3 mL) under a nitrogenatmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-aceticacid (prepared as described in Example 29a, 136 mg, 0.410 mmol),N-methylmorpholine (87.1 mg, 0.861 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (82.3 mg,0.431 mmol) were added sequentially. After stirring at 25° C. for 90min, 1.0M aqueous hydrochloric acid solution (10 mL) and saturatedaqueous brine solution (10 mL) were added to the reaction mixture. Themixture was extracted with ethyl acetate (3×10 mL) and the combinedorganic layers were dried over sodium sulfate, filtered, andconcentrated in vacuo. The crude product,(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, was used in the next step without furtherpurification. LC-MS calcd for C₂₅H₃₃N₃O₇S₂ 551.18, found 552.3 [M+H⁺].

b)(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide

The crude(rac-di-exo)-3-[[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester was dissolved in ethanol (10 mL). To this solution wasadded a 21 wt. % solution of sodium ethoxide in ethanol (0.399 g, 1.23mmol) and the reaction mixture was stirred at 25° C. for 2.5 h. A 1.0Maqueous hydrochloric acid solution (10 mL) was added and, after stirringfor another 30 min, additional 1.0M aqueous hydrochloric acid solution(5 mL) was added upon which the product precipitated. The solid wascollected by vacuum filtration, washed with 1.0M aqueous hydrochloricacid solution (5 mL), and then dried in vacuo, to afford the desiredproduct,(rac-di-exo)-N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,4]thiazin-7-yl}-methanesulfonamide(48.6 mg, 0.094 mmol, 22.8% over two steps), as a tan solid. ¹H NMR (400MHz, CDCl₃) δ: 0.96-1.01 (6H, m), 1.26-1.29 (2H, m), 1.48-1.73 (5H, m),2.58-2.63 (1H, m), 3.07 (3H, s), 3.08-3.17 (1H, m), 3.25 (1H, s),3.37-3.40 (1H, m), 3.89-3.97 (1H, m), 4.90 (0.7H, d, J=17.3 Hz), 5.21(0.3H, d, J=17.4 Hz), 5.28 (0.7H, d, J=16.7 Hz), 5.72 (0.3H, d, J=17.1Hz), 6.17-6.22 (1H, m), 6.35-6.39 (1H, m), 7.23 (1H, d, J=8.5 Hz), 7.38(1H, s), 7.56-7.62 (1H, m), 7.68-7.69 (1H, m). LC-MS calcd forC₂₄H₂₉N₃O₆S₂ 519.15, found 520.2 [M+H⁺].

Example 39(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

a)(rac-di-exo)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one

To a stirred solution of(rac-di-exo)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 9b, 400 mg, 1.454mmol) in anhydrous N,N-dimethylformamide (14 mL) under a nitrogenatmosphere,(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,532.1 mg, 1.454 mmol), N-methylmorpholine (309 mg, 3.053 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (292 mg,1.527 mmol) were added sequentially. After stirring at 25° C. for 4 h,triethylamine (441 mg, 4.362 mmol) was added, and the mixture wasstirred at 50° C. for 2 h. The reaction mixture was allowed to cool to25° C. and continued to stir for 16 h. Upon addition of a 1.0M aqueoushydrochloric acid solution (10 mL), a white precipitate formed that wascollected by vacuum filtration, washed with 1.0M aqueous hydrochloricacid solution (5 mL), and dried in vacuo to afford the desired product,(rac-di-exo)-3-(4-fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(280.2 mg, 0.474 mmol, 32.6%), as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.16-1.20 (2H, m), 1.38 (1H, d, J=9.4 Hz), 1.63 (1H, d,J=9.3 Hz), 2.86 (1H, bs), 3.21 (1H, bs), 3.41 (1H, d, J=9.2 Hz), 4.53(1H, d, J=15.6 Hz), 5.05 (1H, d, J=15.6 Hz), 6.13-6.16 (1H, m),6.35-6.37 (1H, m), 7.15 (2H, t, J=8.4 Hz), 7.33-7.38 (3H, m), 7.99-8.01(1H, m), 8.09 (1H, s). LC-MS calcd for C₂₄H₁₉FIN₃O₄S 591.01, found 592.4[M+H⁺].

b)(rac-di-exo)-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

(rac-di-exo)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(0.100 g, 0.1692 mmol), potassium triphosphate (0.1075 g, 0.5076 mmol),sarcosine (N-methyl glycine) (0.0094 g, 0.1015 mmol), and copper (I)iodide (0.00128 g, 0.0677 mmol) were combined. AnhydrousN,N-dimethylformamide (3 mL) was added followed by N-methylmethanesulfonamide (0.09223 g, 0.8460 mmol). The solution was degassedand backfilled with nitrogen. The mixture was stirred at 100° C. for 16h. Upon cooling, the mixture was diluted with ethyl acetate (200 mL),washed with 1.0M aqueous hydrochloric acid solution (2×100 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to a solid.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 100% ethyl acetate in hexanes) afforded the desiredproduct,(rac-di-exo)-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide(0.00105 g, 0.0184 mmol, 10.8%), as a pale yellow solid. ¹H NMR (400MHz, CDCl₃) δ: 1.27 (3H, s), 1.56 (1H, d, J=9.5 Hz), 1.66 (1H, d, J=9.2Hz), 2.19 (1H, s), 2.64 (1H, s), 2.75 (1H, d, J=9.3 Hz), 3.19 (1H, bs),3.39 (1H, d, J=9.4 Hz), 3.44 (1H, bs), 3.75 (1H, s), 4.32 (1H, d, J=15.4Hz), 5.22 (1H, d, J=14.8 Hz), 6.11-6.13 (1H, m), 6.36-6.38 (1H, m), 6.97(1H, d, J=8.6 Hz), 7.04 (2H, t, J=8.6 Hz), 7.20-7.24 (2H, m), 7.85 (1H,dd, J₁=8.6 Hz, J₂=2.2 Hz), 8.22 (1H, s). LC-MS calcd for C₂₆H₂₅FN₄O₆S₂572.12, found 573.3 [M+H⁺].

Example 40 Cyclopropanesulfonic acid{3-(1R,2S,7R,8S)-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

A reaction flask was charged with copper (I) iodide (20 mg, 0.11 mmol),sarcosine (N-methyl glycine) (14.7 mg, 0.17 mmol),cyclopropanesulfonamide (125 mg, 1.04 mmol),(1R,2S,7R,8S)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 34, 115 mg, 0.21 mmol) and potassiumphosphate (176 mg, 0.83 mmol). The flask was degassed and backfilledwith nitrogen, and then anhydrous N,N-dimethylformamide (5 mL) wasadded. The resulting suspension was vigorously stirred at 100° C. for 3h, and then allowed to cool to 25° C. The mixture was passed through aplug of Celite and rinsed with 10% methanol/dichloromethane. Thefiltrate was concentrated in vacuo, and the residue was purified byprep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron,30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct, cyclopropanesulfonic acid{3-(1R,2S,7R,8S)-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(25 mg, 0.046 mmol, 22%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ:0.92-0.98 (10H, m), 1.20-1.64 (9H, m), 2.52 (1H, m), 2.62 (1H, s), 2.68(1H, m), 3.00 (1H, d, J=9.6 Hz), 3.07 (1H, m), 3.61 (1H, d, J=10.4 Hz),3.66 (1H, m), 7.50-7.59 (3H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₅H₃₂N₄O₆S₂ 548.18, found 549.4 [M+H⁺].

Example 41N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-benzenesulfonamide

(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 0.10 g, 0.17 mmol), cyclopropanesulfonic acid amide (0.21 mg, 1.36 mmol), sarcosine (N-methyl glycine)(0.02 g, 0.26 mmol), copper (I) iodide (0.03 g, 0.17 mmol), andpotassium phosphate (0.22 g, 1.02 mmol) were combined and dissolved inN,N-dimethylformamide (10 mL). The flask was degassed and backfilledwith nitrogen (3×). The reaction was stirred at 100° C. for 4 h. Themixture was allowed to cool to 25° C., diluted with ethyl acetate (20mL), and extracted with saturated aqueous sodium bicarbonate solution(2×20 mL). The combined organic layers were dried over magnesiumsulfate, filtered and concentrated in vacuo. Purification by flashcolumn chromatography (Teledyne Isco RediSep column; 0 to 8% methanol indichloromethane) afforded the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-benzenesulfonamide(0.10 g, 0.16 mmol, 92%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.16-1.19 (3H, m), 1.38-1.57 (5H, m), 2.98 (1H, d, J=8.4 Hz), 3.51(1H, d, J=9.2 Hz), 4.40 (1H, d, J=15.6 Hz), 4.94 (1H, d, J=15.6 Hz),7.11-7.15 (1H, m), 7.29-7.32 (2H, m), 7.39-7.47 (2H, m), 7.54-7.64 (4H,m), 7.74-7.82 (3H, m), 10.74 (1H, s). LC-MS (ESI) calcd forC₃₀H₂₇FN₄O₆S₂ 622.69, found 623.3 [M+H⁺].

Example 42 (rac-di-exo)-Cyclopropanesulfonic acid{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

A reaction flask was charged with copper (I) iodide (20 mg, 0.11 mmol),sarcosine (N-methyl glycine) (14.7 mg, 0.17 mmol),cyclopropanesulfonamide (125 mg, 1.04 mmol),(rac-di-exo)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 33, 115 mg, 0.21 mmol) and potassiumphosphate (175 mg, 0.82 mmol). The flask was degassed and backfilledwith nitrogen, and then anhydrous N,N-dimethylformamide (5 mL) wasadded. The resulting suspension was vigorously stirred at 100° C. for 5h, and then allowed to cool to 25° C. The mixture was passed through aplug of Celite and rinsed with 10% methanol/dichloromethane. Thefiltrate was concentrated in vacuo, and the residue was purified byprep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron,30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct, (rac-di-exo)-cyclopropanesulfonic acid{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(17 mg, 0.031 mmol, 15%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ:0.91-0.96 (10H, m), 1.40-1.76 (7H, m), 2.69 (1H, m), 3.00 (1H, bs), 3.29(1H, m), 3.84 (2H, m), 4.74 (2H, m), 7.51-7.59 (3H, m), 10.17 (1H, s).LC-MS (ESI) calcd for C₂₄H₃₀N₄O₇S₂ 550.16, found 551.4 [M+H⁺].

Example 43 (rac-di-exo)-Cyclopropanesulfonic acid{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

a)(rac-di-exo)-3-{(3,3-Dimethyl-butyl)-[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a solution of(rac-di-exo)-3-(3,3-dimethyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 32a, 227.8 mg, 0.907mmol) in N,N-dimethylformamide (3 mL) was added(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,398.2 mg, 1.088 mmol) and the mixture was vortexed until all materialhad completely dissolved. A 1.0M solution ofN,N-dicyclohexylcarbodiimide in dichloromethane (1.179 mL, 1.179 mmol)was added to the above solution and the mixture was stirred at 25° C.for 18 h under a nitrogen atmosphere. The precipitatedN,N-dicyclohexylurea was removed by filtration over Celite, the filtercake was washed with dichloromethane (3×5 mL), and the solvent wasremoved in vacuo. The residue was dissolved in a 1:1 mixture of ethylacetate and hexanes (20 mL), washed with saturated aqueous brinesolution, saturated aqueous sodium bicarbonate solution and water. Thelayers were separated and the aqueous layers were back-extracted with a1:1 mixture of ethyl acetate and hexanes (20 mL). The combined organiclayers were dried over sodium sulfate, filtered, and concentrated invacuo to afford the crude product,(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, as a yellow oil, which was used in the next stepwithout any further purification. LC-MS calcd for C₂₄H₃₀₁N₃O₅S 599.10,found 600.3 [M+H⁺].

b)(rac-di-exo)-3-(3,3-Dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one

The crude(rac-di-exo)-3-{(3,3-dimethyl-butyl)-[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (877.2 mg, 0.907 mmol) was dissolved in ethanol (20mL). To this solution was added a 21 wt. % solution of sodium ethoxidein ethanol (1.176 g, 3.628 mmol) and the reaction mixture was stirred at25° C. for 2 h. A 1.0M aqueous hydrochloric acid solution (20 mL) wasadded upon which the product began to precipitate. The reaction mixturewas stirred for another 30 min before the precipitate was collected byvacuum filtration. The solid was washed with 1.0M aqueous hydrochloricacid solution followed by water and was then dried in vacuo. Theresulting solid was then triturated with methanol, filtered and dried invacuo. Further purification by flash column chromatography (TeledyneIsco RediSep column; 0 to 50% ethyl acetate in hexanes) afforded thedesired product,(rac-di-exo)-3-(3,3-dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(154.1 mg, 0.272 mmol, 30%), as a white solid. ¹H NMR (400 MHz, CDCl₃)δ: 1.01 (9H, s), 1.32-2.00 (4H, m), 2.19 (1H, d, J=2.2 Hz), 2.75 (1H, d,J=9.5 Hz), 3.10-3.52 (4H, m), 3.78-3.88 (1H, m), 6.17-6.20 (1H, m),6.40-6.46 (1H, m), 7.01 (1H, d, J=8.6 Hz), 7.84 (1H, dd, J₁=8.5 Hz,J₂=1.5 Hz), 8.22 (1H, s). LC-MS calcd for C₂₃H₂₆₁N₃O₄S 567.07, found568.3 [M+H⁺].

c) (rac-di-exo)-Cyclopropanesulfonic acid{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

(rac-di-exo)-3-(3,3-Dimethyl-butyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(0.155 g, 0.255 mmol), potassium triphosphate (0.271 g, 1.275 mmol),sarcosine (N-methyl glycine) (0.027 g, 0.306 mmol), and copper (I)iodide (0.024 g, 0.128 mmol) were combined. AnhydrousN,N-dimethylformamide (7 mL) was added followed by cyclopropanesulfonicacid amide (0.155 g, 1.275 mmol). The solution was degassed andbackfilled with nitrogen. The mixture was stirred at 100° C. for 16 h.Upon cooling, the mixture was diluted with ethyl acetate (200 mL),washed with 1.0M aqueous hydrochloric acid solution (2×100 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to a solid.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 100% ethyl acetate in hexanes) afforded the desiredproduct, (rac-di-exo)-cyclopropanesulfonic acid{3-[3-(3,3-dimethyl-butyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(0.089 g, 0.159 mmol, 62.5%), as a pale yellow solid. ¹H NMR (400 MHz,CDCl₃) δ: 1.00 (9H, s), 1.14-1.21 (2H, m), 1.24-1.37 (2H, m), 1.51-1.63(2H, m), 2.49-2.61 (1H, m), 2.73-2.76 (1H, m), 3.09-3.16 (1H, m),3.19-3.25 (1H, m), 3.38-3.46 (2H, m), 3.79-3.88 (1H, m), 6.17-6.19 (1H,m), 6.38-6.40 (1H, m), 7.18-7.25 (1H, m), 7.57 (1H, s), 7.63-7.66 (1H,m), 7.73-7.79 (1H, m). LC-MS (ESI) calcd for C₂₆H₃₂N₄O₆S₂ 560.18, found561.6 [M+H⁺].

Example 44N-[3-(1R,2S,7R,8S)-3-Cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

a) (1S,2R,3S,4R)-3-Cyclopentylamino-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

Cyclopentanone (0.12 mL, 1.38 mmol) was added to a solution of(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 230 mg, 1.26 mmol) inanhydrous methanol (10 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 10 min, glacial acetic acid (0.5 mL) and sodiumcyanoborohydride (260 mg, 3.15 mmol) were added sequentially, and theresulting mixture was stirred at 50° C. for 30 min. The reaction mixturewas poured into saturated aqueous sodium bicarbonate solution andextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over sodium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct,(1S,2R,3S,4R)-3-cyclopentylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester (237 mg, 0.94 mmol, 75%), as a yellow oil. LC-MS (ESI) calcdfor C₁₅H₂₅NO₂ 251.19, found 252.0 [M+H⁺].

b)N-[3-(1R,2S,7R,8S)-3-Cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

To a stirred solution of(1S,2R,3S,4R)-3-cyclopentylamino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (150 mg, 0.60 mmol) and(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 181 mg, 0.54 mmol) inanhydrous N,N-dimethylformamide (5 mL) under a nitrogen atmosphere,N-methylmorpholine (0.12 mL, 1.08 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (104 mg,0.54 mmol) were added sequentially. The mixture was stirred at 25° C.for 45 min, triethylamine (0.25 mL, 1.76 mmol) was added and theresulting mixture was stirred at 50° C. for 60 h. The reaction mixturewas allowed to cool to 25° C., diluted with ethyl acetate, washed with1.0M aqueous hydrochloric acid solution and saturated aqueous brinesolution, dried over magnesium sulfate and filtered. The filtrate wasconcentrated in vacuo and the residue was purified by prep-HPLC [ColumnLuna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron, 30%-95% in 7 min @30mL/min flow rate, 0.05% trifluoroacetic acid in acetonitrile/0.05%trifluoroacetic acid in water] to afford the desired product,N-[3-(1R,2S,7R,8S)-3-cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(80 mg, 0.15 mmol, 26%), as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ:1.20-1.65 (8H, m), 1.75-1.95 (6H, m), 2.42 (1H, s), 2.60 (1H, s), 2.99(1H, d, J=9.2 Hz), 3.05 (3H, s), 3.60 (1H, d, J=9.2 Hz), 3.93 (1H, m),7.48-7.58 (3H, m), 10.17 (1H, s). LC-MS (ESI) calcd for C₂₃H₂₈N₄O₆S₂520.15, found 521.4 [M+H⁺].

Example 45 (rac-di-exo)-Cyclopropanesulfonic acid{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

a)(rac-di-exo)-3-[[2-(7-Iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester

To a solution of(rac-di-exo)-3-(3-methyl-butylamino)-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester (prepared as described in Example 31b, 214.8 mg, 0.906mmol) in N,N-dimethylformamide (3 mL) was added(7-iodo-1,1-dioxo-1,4-dihydro-1)⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in US patent application US 2008/0031852,397.7 mg, 1.087 mmol) and the mixture was vortexed until all materialhad completely dissolved. A 1.0M solution ofN,N-dicyclohexylcarbodiimide in dichloromethane (1.178 mL, 1.178 mmol)was added to the above solution and the mixture was stirred at 25° C.for 18 h under a nitrogen atmosphere. The precipitatedN,N-dicyclohexylurea was removed by filtration over Celite, the filtercake was washed with dichloromethane (3×5 mL), and the solvent wasremoved in vacuo. The residue was dissolved in a 1:1 mixture of ethylacetate and hexanes (20 mL), washed with saturated aqueous brinesolution, saturated aqueous sodium bicarbonate solution and water. Thelayers were separated and the aqueous layers were back-extracted with a1:1 mixture of ethyl acetate and hexanes (20 mL). The combined organiclayers were dried over sodium sulfate, filtered, and concentrated invacuo to afford the crude product,(rac-di-exo)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester, as a yellow oil, which was used in the next stepwithout any further purification. LC-MS calcd for C₂₃H₂₈₁N₃O₅S 585.08,found 586.3 [M+H⁺].

b)(rac-di-exo)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1)⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one

The crude(rac-di-exo)-3-[[2-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-(3-methyl-butyl)-amino]-bicyclo[2.2.1]hept-5-ene-2-carboxylicacid methyl ester was dissolved in ethanol (20 mL). To this solution wasadded a 21 wt. % solution of sodium ethoxide in ethanol (1.174 g, 3.624mmol) and the reaction mixture was stirred at 25° C. for 2 h. A 1.0Maqueous hydrochloric acid solution (20 mL) was added upon which theproduct began to precipitate. The reaction mixture was stirred foranother 30 min before the precipitate was collected by vacuumfiltration. The solid was washed with 1.0M aqueous hydrochloric acidsolution followed by water and was then dried in vacuo. The resultingsolid was then triturated with methanol, filtered and dried in vacuo.Further purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 50% ethyl acetate in hexanes) afforded the desiredproduct,(rac-di-exo)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(28.7 mg, 0.0519 mmol, 5.7% over two steps), as an off-white solid. ¹HNMR (400 MHz, CDCl₃) δ: 0.98 (3H, d, J=3.3 Hz), 0.99 (3H, d, J=2.9 Hz),1.47-1.68 (6H, m), 2.75 (1H, d, J=9.3 Hz), 3.08-3.32 (2H, m), 3.39-3.44(2H, m), 3.82-3.89 (1H, m), 6.18-6.20 (1H, m), 6.39-6.41 (1H, m), 6.99(1H, d, J=8.6 Hz), 7.84 (1H, d, J=10.8 Hz), 8.21 (1H, s). LC-MS calcdfor C₂₂H₂₄₁N₃O₄S 553.05, found 554.1 [M+H⁺].

c) (rac-di-exo)-Cyclopropanesulfonic acid{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

(rac-di-exo)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(0.029 g, 0.052 mmol), potassium triphosphate (0.055 g, 0.259 mmol),sarcosine (N-methyl glycine) (0.006 g, 0.062 mmol), and copper (I)iodide (0.005 g, 0.026 mmol) were combined. AnhydrousN,N-dimethylformamide (7 mL) was added followed by cyclopropanesulfonicacid amide (0.031 g, 0.259 mmol). The solution was degassed andbackfilled with nitrogen. The mixture was stirred at 100° C. for 16 h.Upon cooling, the mixture was diluted with ethyl acetate (200 mL),washed with 1.0M aqueous hydrochloric acid solution (2×100 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to a solid.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 100% ethyl acetate in hexanes) afforded the desiredproduct, (rac-di-exo)-cyclopropanesulfonic acid{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(0.009 g, 0.016 mmol, 31.6%), as a pale yellow solid. ¹H NMR (400 MHz,CDCl₃) δ: 0.98-1.08 (6H, m), 1.17-1.26 (3H, m), 1.52-1.73 (4H, m),2.48-2.62 (2H, m), 2.75-2.77 (1H, m), 3.08-3.15 (1H, m), 3.39-3.44 (2H,m), 3.83-3.91 (1H, m), 6.19 (1H, dd, J₁=5.5 Hz, J₂=3.1 Hz), 6.40 (1H,dd, J₁=5.4 Hz, J₂=4.0 Hz), 7.01 (1H, s), 7.24 (1H, t, J=7.1 Hz),7.63-7.65 (1H, m), 7.70-7.73 (1H, m). LC-MS (ESI) calcd for C₂₅H₃₀N₄O₆S₂546.16, found 547.4 [M+H⁺].

Example 46 (rac-di-exo)-Cyclopropanesulfonic acid{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

(rac-di-exo)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-4-one(prepared as described in Example 39a, 0.169 g, 0.285 mmol), potassiumtriphosphate (0.303 g, 1.426 mmol), sarcosine (N-methyl glycine) (0.031g, 0.342 mmol), and copper (I) iodide (0.027 g, 0.146 mmol) werecombined. Anhydrous N,N-dimethylformamide (7 mL) was added followed bycyclopropanesulfonic acid amide (0.173 g, 1.426 mmol). The solution wasdegassed and backfilled with nitrogen. The mixture was stirred at 100°C. for 16 h. Upon cooling, the mixture was diluted with ethyl acetate(200 mL), washed with 1.0M aqueous hydrochloric acid solution (2×100mL), dried over magnesium sulfate, filtered, and concentrated in vacuoto a solid. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 100% ethyl acetate in hexanes) afforded the desiredproduct, (rac-di-exo)-cyclopropanesulfonic acid{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undeca-5,9-dien-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(0.093 g, 0.159 mmol, 55.8%), as a pale yellow solid. ¹H NMR (400 MHz,CDCl₃) δ: 0.97-1.04 (2H, m), 1.14-1.20 (2H, m), 1.53 (1H, d, J=9.4 Hz),1.66 (1H, d, J=9.5 Hz), 2.48-2.61 (2H, m), 2.74 (1H, d, J=9.4 Hz),3.37-3.41 (2H, m), 4.32 (1H, d, J=15.3 Hz), 5.19 (1H, d, J=14.8 Hz),6.09-6.11 (1H, m), 6.33-6.36 (1H, m), 7.02 (2H, t, J=8.6 Hz), 7.16-7.23(3H, m), 7.63 (1H, dd, J₁=9.3 Hz, J₂=2.3 Hz), 7.77 (1H, d, J=2.3 Hz),8.04 (1H, s). LC-MS (ESI) calcd for C₂₇H₂₅FN₄O₆S₂ 584.12, found 585.2[M+H⁺].

Example 47N-[3-(1R,2S,7R,8S)-(3-Cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide-N-isopropylcarbamate

N-[3-(1R,2S,7R,8S)-3-cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(prepared as described in Example 44, 48.5 mg, 0.093 mmol) was dissolvedin anhydrous pyridine (2 mL) under a nitrogen atmosphere, a 1.0Msolution of isopropyl chloroformate in toluene (0.46 mL, 0.46 mmol) wasadded slowly. The resulting mixture was stirred at 25° C. for 15 min,LC-MS showed the completion of the reaction. The reaction was quenchedwith methanol (1 mL) and the resulting mixture was concentrated invacuo. The residue was purified by prep-HPLC [Column Luna 5μ C18 (2) 100Å AXIA 150×21.2 mm, 5 micron, 30%-95% in 7 min @30 mL/min flow rate,0.05% trifluoroacetic acid in acetonitrile/0.05% trifluoroacetic acid inwater] to afford the desired product,N-[3-(1R,2S,7R,8S)-(3-cyclopentyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide-N-isopropylcarbamate (15 mg, 0.025 mmol, 27%), as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.19 (6H, d, J=6.0 Hz), 1.30-1.65 (8H, m), 1.75-1.95 (6H,m), 2.43 (1H, m), 2.61 (1H, m), 3.00 (1H, m), 3.55 (1H, m), 3.62 (3H,s), 3.95 (1H, m), 4.94 (1H, m), 7.57 (1H, d, J=9.2 Hz), 7.64 (1H, dd,J=8.8, 1.6 Hz), 7.97 (1H, s). LC-MS (ESI) calcd for C₂₇H₃₄N₄O₈S₂ 606.16.found 607.3 [M+H⁺].

Example 48(rac-di-exo)-N-[3-(3-Cyclopentyl-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

a)(rac-di-exo)-3-Cyclopentylamino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester

Cyclopentanone (0.56 mL, 6.37 mmol) was added to a solution of(rac-di-exo)-3-amino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylic acidmethyl ester (prepared as described in Example 8a, 1.10 g, 6.37 mmol) inanhydrous methanol (15 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 20 min, glacial acetic acid (0.75 mL) and sodiumcyanoborohydride (1.0 g, 15.92 mmol) were added sequentially, and theresulting mixture was stirred at 50° C. for 1 h. The reaction mixturewas poured into saturated aqueous sodium bicarbonate solution andextracted with ethyl acetate. The combined organic layers were washedwith saturated aqueous brine solution, dried over sodium sulfate andfiltered. The filtrate was concentrated in vacuo to afford the desiredproduct,(rac-di-exo)-3-cyclopentylamino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (1.06 g, 4.43 mmol, 70%), as a yellow oil. LC-MS (ESI)calcd for C₁₃H₂₁NO₃ 239.15, found 240.2 [M+H⁺].

b)(rac-di-exo)-N-[3-(3-Cyclopentyl-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

To a stirred solution of(rac-di-exo)-3-cyclopentylamino-7-oxa-bicyclo[2.2.1]heptane-2-carboxylicacid methyl ester (170 mg, 0.71 mmol) and(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 197 mg, 0.60 mmol) inanhydrous N,N-dimethylformamide (5 mL) under a nitrogen atmosphere,N-methylmorpholine (0.132 mL, 1.20 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (115 mg,0.60 mmol) were added sequentially. The mixture was stirred at 25° C.for 1 h, triethylamine (0.34 mL, 2.4 mmol) was added and the resultingmixture was stirred at 50° C. for 1 h. The reaction mixture was allowedto cool to 25° C., diluted with ethyl acetate, washed with 1.0M aqueoushydrochloric acid solution and saturated aqueous brine solution, driedover magnesium sulfate and filtered. The filtrate was concentrated invacuo and the residue was purified by prep-HPLC [Column Luna 5μ C18 (2)100A AXIA 150×21.2 mm, 5 micron, 30%-95% in 7 min @30 mL/min flow rate,0.05% trifluoroacetic acid in acetonitrile/0.05% trifluoroacetic acid inwater] to afford the desired product,(rac-di-exo)-N-[3-(3-cyclopentyl-6-hydroxy-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(120 mg, 0.23 mmol, 38%), as an off-white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.48-1.96 (12H, m), 3.05 (3H, s), 3.30 (1H, d, J=8.0 Hz),3.89 (1H, d, J=9.2 Hz), 4.13 (1H, m), 4.57 (1H, s), 4.74 (1H, s),7.49-7.58 (3H, m), 10.17 (1H, s). LC-MS (ESI) calcd for C₂₂H₂₆N₄O₇S₂522.12, found 523.4 [M+H⁺].

Example 49N-{3-[(2S,7R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) 4-Oxa-tricyclo[5.2.2.0^(2,6)]undecane-3,5-dione

4-Oxa-tricyclo[5.2.2.0^(2,6)]undec-8-ene-3,5-dione (4.00 g, 22.45 mmol)was dissolved in ethyl acetate (100 mL). 10% Palladium on carbon (400mg) was added. The flask was degassed and backfilled with hydrogen gasvia balloon. The mixture was stirred at 25° C. for 16 h. The mixture waspassed through a plug of Celite and the filtrate was concentrated invacuo to afford a thick clear oil. Purification by flash columnchromatography (Teledyne Isco RediSep column; 0 to 30% ethyl acetate inhexanes) afforded the desired product,4-oxa-tricyclo[5.2.2.0^(2,6)]undecane-3,5-dione (2.92 g, 16.20 mmol,72%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆) δ: 1.55-1.64 (6H, m),1.76 (2H, d, J=9.2 Hz), 2.25 (2H, s), 3.11 (2H, s). LC-MS (ESI) calcdfor C₁₀H₁₂O₃ 180.20, found 181.0 [M+H⁺].

b) (2S,3R)-Bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester

4-Oxa-tricyclo[5.2.2.0^(2,6)]undec-3,5-dione (0.90 g, 4.99 mmol) wasdissolved in toluene (50 mL) and carbon tetrachloride (50 mL). Quinine(1.78 g, 5.49 mmol) was added and the mixture was cooled to −55° C.Methanol (0.61 mL, 14.97 mmol) was added dropwise to the above mixture.The reaction was stirred at −55° C. for 18 h. The reaction was warmed to25° C. and concentrated in vacuo. The crude material was dissolved inethyl acetate (50 mL) and washed with 1.0M aqueous hydrochloric acidsolution (2×40 mL). The organic layer was further washed with saturatedaqueous brine solution (20 mL), dried over magnesium sulfate, filtered,and concentrated in vacuo to afford a clear oil. Purification by flashcolumn chromatography (Teledyne Isco RediSep column; 0 to 50% ethylacetate in hexanes) afforded the desired product,(2S,3R)-bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester(1.10 g, 5.18 mmol, 92%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆) δ:1.31 (2H, dd, J₁=20.0 Hz, J₂=12.4 Hz), 1.52-1.54 (4H, m), 1.63 (1H, t,J=10.4 Hz), 1.75 (1H, t, J=9.6 Hz), 1.87 (2H, bs), 2.84 (2H, dd, J₁=29.6Hz, J₂=10.8 Hz), 3.52 (3H, s), 12.01 (1H, s). LC-MS (ESI) calcd forC₁₁H₁₆O₄ 212.24, found 213.1 [M+H⁺].

c) (2R,3S)-3-Benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylicacid methyl ester

(2S,3R)-Bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester(1.01 g, 4.76 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL).The flask was degassed and backfilled with nitrogen and the mixture wascooled to 0° C. Triethylamine (1.99 mL, 14.28 mmol) was added followedby the dropwise addition of ethyl chloroformate (0.91 mL, 9.52 mmol)with vigorous stirring. The mixture was stirred at 0° C. for 1 h. Sodiumazide (0.93 g, 14.28 mmol) was dissolved in water (5 mL) and added tothe reaction mixture at 0° C. The mixture was stirred at 0° C. for 5min. The ice bath was removed. The mixture was warmed to 25° C. and wasstirred for 2 h. The mixture was poured into water (50 mL) and theproduct extracted into ethyl acetate (50 mL). The organic layer wasfurther washed with half-saturated aqueous sodium bicarbonate solution(2×20 mL), saturated aqueous brine solution (20 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. The oil was dissolved in anhydrous benzene (10 mL) and refluxedwhile stirring under nitrogen for 2 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a slightly yellow oil. Theoil was dissolved in dichloromethane (10 mL) and benzyl alcohol (0.54mL, 5.24 mmol) was added followed by triethylamine (1.33 mL, 9.52 mmol).The mixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C.the solution was concentrated in vacuo to afford a golden oil.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 20% ethyl acetate in hexanes) afforded the desired product,(2R,3S)-3-benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester (0.58 g, 1.83 mmol, 38%), as a clear oil. ¹H NMR (400 MHz,CDCl₃) δ: 1.18-1.28 (2H, m), 1.42-1.50 (5H, m), 1.73-1.96 (3H, m), 2.88(1H, d, J₁=5.6 Hz), 3.27 (1H, s), 3.42 (3H, s), 4.00-4.04 (1H, m), 4.97(2H, dd, J₁=46.4 Hz, J₂=12.8 Hz), 7.06 (1H, d, J=9.6 Hz), 7.24-7.34 (4H,m). LC-MS (ESI) calcd for C₁₈H₂₃NO₄ 317.38, found 317.9 [M+H⁺].

d) (2R,3S)-3-Amino-bicyclo[2.2.2]octane-2-carboxylic acid methyl esterhydrochloride

(2R,3S)-3-Benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester (0.57 g, 1.79 mmol) was dissolved in ethyl acetate (20 mL).10% Palladium on carbon (60 mg) was added. The flask was degassed andbackfilled with hydrogen gas via balloon. The mixture was stirred at 25°C. for 16 h. The mixture was passed through a plug of Celite and thefiltrate was concentrated in vacuo to afford a thick clear oil. The oilwas dissolved in diethyl ether (6 mL) and added dropwise with vigorousstirring to a mixture of 4.0M solution of hydrochloric acid in1,4-dioxane (1.02 mL) and diethyl ether (10 mL). The desired productbegan to precipitate as a white solid. The mixture was stirred for 20min. The precipitate was collected by vacuum filtration, washed withadditional diethyl ether (5 mL). The solid was further dried in vacuofor 1 h to afford the desired product,(2R,3S)-3-amino-bicyclo[2,2,2]octane-2-carboxylic acid methyl esterhydrochloride (0.33 g, 1.50 mmol, 84%), as a white powder. ¹H NMR (400MHz, DMSO-d₆) δ: 1.38 (2H, dd, J₁=21.2 Hz, J₂=13.6 Hz), 1.55-1.63 (5H,m), 1.76-1.89 (3H, m), 3.02 (1H, dd, J₁=10.0 Hz, J₂=2.4 Hz), 3.47 (1H,bs), 3.65 (3H, s), 7.97 (3H, s). LC-MS (ESI) calcd for C₁₀H₁₇NO₂ (freeamine) 183.25, found 184.2 [M+H⁺].

e) (2R,3S)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.2]octane-2-carboxylicacid methyl ester

(2R,3S)-3-Amino-bicyclo[2,2,2]octane-2-carboxylic acid methyl esterhydrochloride (0.34 g, 1.54 mmol) was dissolved in methanol (10 mL).Sodium acetate (0.25 g, 3.08 mmol) was added followed by 4 Å powderedmolecular sieves (0.34 g) and 4-fluoro-benzaldehyde (0.16 mL, 1.54mmol). Sodium cyanoborohydride (0.19 g, 3.08 mmol) was added and themixture was stirred at 25° C. for 16 h. The mixture was poured into amixture of saturated aqueous sodium bicarbonate solution (20 mL) andethyl acetate (30 mL). After shaking, both layers were passed through aplug of Celite. The organic layer was further washed with saturatedaqueous sodium bicarbonate solution (10 mL), saturated aqueous brinesolution (10 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product,(2R,3S)-3-(4-fluoro-benzylamino)-bicyclo[2,2,2]octane-2-carboxylic acidmethyl ester (0.32 g, 1.11 mmol, 72%), as a clear oil. LC-MS (ESI) calcdfor C₁₇H₂₂FNO₂ 291.36, found 292.2 [M+H⁺].

f)N-{3-[(2S,7R)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(2R,3S)-3-(4-Fluoro-benzylamino)-bicyclo[2,2,2]octane-2-carboxylic acidmethyl ester (93 mg, 0.32 mmol) was dissolved in anhydrousN,N-dimethylformamide (4 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 107 mg, 0.32 mmol) was addedfollowed by N-methylmorpholine (74 μL, 0.67 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (65 mg, 0.34mmol) was added and the mixture was stirred at 25° C. for 16 h. Thereaction was quenched via addition of saturated aqueous sodiumbicarbonate solution (20 mL). The mixture was extracted with ethylacetate (3×30 mL). The combined organic layers were washed withsaturated aqueous brine solution (20 mL), dried over magnesium sulfate,filtered, and concentrated in vacuo to afford a golden oil. The oil wasdissolved in ethanol (5 mL). A 21 wt. % solution of sodium ethoxide inethanol (0.36 mL, 0.96 mmol) was added. The reaction was refluxed for 16h. The reaction was quenched via the addition of 1.0M aqueoushydrochloric acid solution (10 mL). The mixture was extracted with ethylacetate (3×20 mL). The organic layer was further washed with saturatedsodium bicarbonate solution (2×20 mL), saturated aqueous brine solution(20 mL), dried over magnesium sulfate, filtered, and concentrated invacuo to afford a clear oil. Purification by flash column chromatography(Teledyne Isco RediSep column; 0 to 20% ethyl acetate indichloromethane) afforded the desired product,N-{3-[(2S,7R)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.11 g, 0.19 mmol, 59%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.39 (2H, d, J=8.0 Hz), 1.54-1.59 (8H, m), 1.91 (1H, s), 2.14 (1H,s), 3.06 (3H, s), 3.75 (1H, d, J=11.6 Hz), 4.28 (1H, d, J=15.2 Hz), 5.03(1H, d, J=15.6 Hz), 7.13-7.17 (2H, m), 7.34-7.37 (2H, m), 7.50-7.60 (3H,m), 10.18 (1H, s). LC-MS (ESI) calcd for C₂₆H₂₇FN₄O₆S₂ 574.64, found575.1 [M+H⁺]. m.p.: 203.8-205.7° C. ee=94.4% [HPLC-analysis: ChiralpakAS-RH 4.6×250 mm, 5 micron, 0.8 mL/min, 310 nm].

Example 50cis-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a) cis-Bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester

4-Oxa-tricyclo[5.2.2.0^(2,6)]undec-8-ene-3,5-dione (1.00 g, 5.61 mmol)was dissolved in methanol (20 mL). 10% Palladium on carbon (100 mg) wasadded. The flask was degassed and backfilled with hydrogen gas viaballoon. The mixture was stirred at 25° C. for 16 h. The mixture waspassed through a plug of Celite and the filtrate was left to stand at25° C. for 72 h. The solution was then concentrated in vacuo to afford aclear oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 60% ethyl acetate in hexanes) afforded the desiredproduct, cis-bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester(1.10 g, 5.18 mmol, 92%), as a clear oil. ¹H NMR (400 MHz, DMSO-d₆) δ:1.31 (2H, dd, J₁=20.0 Hz, J₂=12.4 Hz), 1.52-1.54 (4H, m), 1.63 (1H, t,J=10.4 Hz), 1.75 (1H, t, J=9.6 Hz), 1.87 (2H, bs), 2.84 (2H, dd, J₁=29.6Hz, J₂=10.8 Hz), 3.52 (3H, s), 12.01 (1H, s). LC-MS (ESI) calcd forC₁₁H₁₆O₄ 212.24, found 213.2 [M+H⁺].

b) cis-3-Benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester

cis-Bicyclo[2.2.2]octane-2,3-dicarboxylic acid monomethyl ester (1.09 g,5.14 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL). The flaskwas degassed and backfilled with nitrogen and the mixture was cooled to0° C. Triethylamine (2.15 mL, 15.42 mmol) was added followed by thedropwise addition of ethyl chloroformate (0.98 mL, 10.28 mmol) withvigorous stirring. The mixture was stirred at 0° C. for 1 h. Sodiumazide (1.00 g, 15.42 mmol) was dissolved in water (7 mL) and added tothe reaction mixture at 0° C. The mixture was stirred at 0° C. for 5min. The ice bath was removed. The mixture was warmed to 25° C. and wasstirred for 2 h. The mixture was poured into water (70 mL) and theproduct extracted into ethyl acetate (70 mL). The organic layer wasfurther washed with half-saturated aqueous sodium bicarbonate solution(2×30 mL), saturated aqueous brine solution (30 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. The oil was dissolved in anhydrous benzene (10 mL) and refluxedwhile stirring under nitrogen for 2 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a slightly yellow oil. Theoil was dissolved in dichloromethane (7 mL) and benzyl alcohol (0.58 mL,5.65 mmol) was added followed by triethylamine (1.43 mL, 10.28 mmol).The mixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C.the solution was concentrated in vacuo to afford a golden oil.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 20% ethyl acetate in hexanes) afforded the desired product,cis-3-benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester (1.32 g, 4.16 mmol, 81%), as a clear oil. ¹H NMR (400 MHz,CDCl₃) δ: 1.18-1.28 (2H, m), 1.42-1.50 (5H, m), 1.73-1.96 (3H, m), 2.88(1H, d, J₁=5.6 Hz), 3.27 (1H, s), 3.42 (3H, s), 4.00-4.04 (1H, m), 4.97(2H, dd, J₁=46.4 Hz, J₂=12.8 Hz), 7.06 (1H, d, J=9.6 Hz), 7.24-7.34 (4H,m). LC-MS (ESI) calcd for C₁₈H₂₃NO₄ 317.38, found 317.92 [M+H⁺].

c) cis-3-Amino-bicyclo[2.2.2]octane-2-carboxylic acid methyl esterhydrochloride

cis-3-Benzyloxycarbonylamino-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester (0.67 g, 2.11 mmol) was dissolved in ethyl acetate (10 mL).5% Palladium on carbon (97 mg) was added. The flask was degassed andbackfilled with hydrogen gas via balloon. The mixture was stirred at 25°C. for 16 h. The mixture was passed through a plug of Celite and thefiltrate was concentrated in vacuo to afford a thick clear oil. The oilwas dissolved in diethyl ether (6.7 mL) and added dropwise, withvigorous stirring, to a mixture of 4.0M solution of hydrochloric acid in1,4-dioxane (1.2 mL) in diethyl ether (12 mL). The desired product beganto precipitate as a white solid. The mixture was stirred for 20 min. Theprecipitate was collected by vacuum filtration, washed with additionaldiethyl ether (5 mL). The solid was further dried in vacuo for 1 h toafford the desired product,cis-3-amino-bicyclo[2,2,2]octane-2-carboxylic acid methyl esterhydrochloride (0.35 g, 1.59 mmol, 75%), as a white powder. ¹H NMR (400MHz, DMSO-d₆) δ: 1.38 (2H, dd, J₁=21.2 Hz, J₂=13.6 Hz), 1.55-1.63 (5H,m), 1.76-1.89 (3H, m), 3.02 (1H, dd, J₁=10.0 Hz, J₂=2.4 Hz), 3.47 (1H,bs), 3.65 (3H, s), 7.97 (3H, s). LC-MS (ESI) calcd for C₁₀H₁₇NO₂ (freeamine) 183.25, found 184.05 [M+H⁺].

d) cis-3-(4-Fluoro-benzylamino)-bicyclo[2.2.2]octane-2-carboxylic acidmethyl ester

cis-3-Amino-bicyclo[2,2,2]octane-2-carboxylic acid methyl esterhydrochloride (0.34 g, 1.55 mmol) was dissolved in methanol (10 mL).Sodium acetate (0.25 g, 3.10 mmol) was added followed by 4 Å powderedmolecular sieves (0.34 g) and 4-fluoro-benzaldehyde (0.17 mL, 1.55mmol). Sodium cyanoborohydride (0.20 g, 3.10 mmol) was added and themixture was stirred at 25° C. for 16 h. The mixture was poured into amixture of saturated aqueous sodium bicarbonate solution (20 mL) andethyl acetate (30 mL). After shaking, both layers were passed through aplug of Celite. The organic layer was further washed with saturatedaqueous sodium bicarbonate solution (10 mL), saturated aqueous brinesolution (10 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product,cis-3-(4-fluoro-benzylamino)-bicyclo[2,2,2]octane-2-carboxylic acidmethyl ester (0.34 g, 1.17 mmol, 75%), as a clear oil. LC-MS (ESI) calcdfor C₁₇H₂₂FNO₂ 291.36, found 292.18 [M+H⁺].

d)cis-N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

cis-3-(4-Fluoro-benzylamino)-bicyclo[2,2,2]octane-2-carboxylic acidmethyl ester (0.20 g, 0.69 mmol) was dissolved in anhydrousN,N-dimethylformamide (8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.23 g, 0.69 mmol) was addedfollowed by N-methylmorpholine (0.16 mL, 1.45 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.14 g,0.72 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.29 mL, 2.07 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (50 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×50 mL), saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 20% ethyl acetate in dichloromethane) afforded thedesired product,cis-N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.2.0^(2,7)]dodec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.30 g, 0.52 mmol, 76%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.39 (2H, d, J=8.0 Hz), 1.54-1.59 (8H, m), 1.91 (1H, s), 2.14 (1H,s), 3.06 (3H, s), 3.75 (1H, d, J=11.6 Hz), 4.28 (1H, d, J=15.2 Hz), 5.03(1H, d, J=15.6 Hz), 7.13-7.17 (2H, m), 7.34-7.37 (2H, m), 7.50-7.60 (3H,m), 10.18 (1H, s). LC-MS (ESI) calcd for C₂₆H₂₇FN₄O₆S₂ 574.64, found575.4 [M+H⁺]. m.p.: 203.8-205.7° C. Anal. calcd for C₂₆H₂₇FN₄O₆S₂.0.4PhMe: C, 56.57; H, 4.98; N, 9.16; found C, 57.09; H, 5.08; N, 9.38.

Example 51rac-N-{3-[(1R,2R,7S,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]tridec-5-en-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide

a)rac-(1R,2R,6S,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dioneandrac-(1R,2S,6R,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dione

Furan-2,5-dione (5.32 g, 54.26 mmol) was partially dissolved in xylene(100 mL). Cyclohepta-1,3,5-triene (5 g, 54.26 mmol) was added. Thereaction was stirred at 144° C. for 5 h. The mixture was allowed to coolto 25° C. and concentrated in vacuo. Purification by flash columnchromatography (Teledyne Isco RediSep column; 0 to 20% ethyl acetate inhexanes) afforded the desired products,rac-(1R,2R,6S,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dione(7.46 g, 39.22 mmol, 72%) andrac-(1R,2S,6R,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dione(0.88 g, 4.63 mmol, 8%), as white solids. ¹H NMR (400 MHz, CDCl₃) 8:exo: 0.27-0.42 (2H, m), 1.11-1.15 (2H, m), 3.25 (2H, t, J=1.6 Hz),3.46-3.50 (2H, m), 5.89 (2H, dd, J₁=4.8 Hz, J₂=3.2 Hz); endo: 0.14-0.26(2H, m), 1.12-1.16 (2H, m), 3.09 (2H, s), 3.40-3.44 (2H, m), 5.94 (2H,dd, J₁=4.8 Hz, J₂=3.2 Hz). LC-MS (ESI) calcd for C₁₁H₁₀O₃ 190.20, foundexo 191.0; endo 191.3 [M+H⁺].

b)rac-(1S,2S,4R,5R,6R,7S)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid

rac-(1R,2R,6S,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dione(1.00 g, 5.26 mmol) was dissolved in methanol (20 mL). The reaction wasstirred at 25° C. for 72 h. The mixture was concentrated in vacuo toafford a clear oil. Purification by flash column chromatography(Teledyne Isco RediSep column; 0 to 60% ethyl acetate in hexanes)afforded the desired product,rac-(1S,2S,4R,5R,6R,7S)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid (1.11 g, 5.00 mmol, 95%), as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.01-0.10 (2H, m), 0.93-1.02 (2H, m), 2.98-3.05 (4H, m),3.45 (3H, s), 5.68-5.77 (2H, m), 11.94 (1H, s). LC-MS (ESI) calcd forC₁₂H₁₄O₄ 222.24, found 223.2 [M+H⁺].

c) rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate

rac-(1S,2S,4R,5R,6R,7S)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid (0.78 g, 3.53 mmol) was dissolved in anhydrous tetrahydrofuran (20mL). The flask was degassed and backfilled with nitrogen and the mixturewas cooled to 0° C. Triethylamine (1.48 mL, 10.59 mmol) was addedfollowed by the dropwise addition of ethyl chloroformate (0.67 mL, 7.06mmol) with vigorous stirring. The mixture was stirred at 0° C. for 1 h.Sodium azide (0.69 g, 10.59 mmol) was dissolved in water (5 mL) andadded to the reaction mixture at 0° C. The mixture was stirred at 0° C.for 5 min. The ice bath was removed. The mixture was warmed to 25° C.and was stirred for 2 h. The mixture was poured into water (70 mL) andthe product extracted into ethyl acetate (70 mL). The organic layer wasfurther washed with half-saturated aqueous sodium bicarbonate solution(2×30 mL), saturated aqueous brine solution (30 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. The oil was dissolved in anhydrous benzene (10 mL) and refluxedwhile stirring under nitrogen for 2 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a slightly yellow oil. Theoil was dissolved in dichloromethane (10 mL) and benzyl alcohol (0.40mL, 3.88 mmol) was added followed by triethylamine (0.98 mL, 7.06 mmol).The mixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C.the solution was concentrated in vacuo to afford a golden oil.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 20% ethyl acetate in hexanes) afforded the desired product,rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate(0.62 g, 1.89 mmol, 54%) as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:0.17-0.22 (2H, m), 0.90-1.05 (2H, m), 2.98-3.14 (3H, m), 3.48 (3H, s),4.38-4.43 (1H, m), 5.00-5.12 (2H, m), 5.77 (1H, t, J=7.2 Hz), 6.05 (1H,t, J=7.2 Hz), 7.29-7.37 (5H, m). LC-MS (ESI) calcd for C₁₉H₂₁NO₄ 327.37,found 328.3 [M+H⁺].

d) rac-Methyl(1R,2R,4S,5S,6S,7R)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylatehydrochloride

rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate(0.61 g, 1.85 mmol) was dissolved in ethyl acetate (10 mL). 5% Palladiumon carbon (0.10 g) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 16 h.The mixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to afford a thick clear oil. The oil was dissolvedin diethyl ether (6 mL) and added dropwise, with vigorous stirring, to amixture of a 4.0M solution of hydrochloric acid in 1,4-dioxane (0.93 mL)in diethyl ether (11 mL). The mixture was concentrated and dried invacuo to afford the desired product, rac-Methyl(1R,2R,4S,5S,6S,7R)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylatehydrochloride (0.43 g, 1.85 mmol, 100%), as a sticky gum. LC-MS (ESI)calcd for C₁₁H₁₇NO₂ (free amine) 195.26, found 196.5 [M+H⁺].

e) rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate

rac-Methyl(1R,2R,4S,5S,6S,7R)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylatehydrochloride (0.43 g, 1.85 mmol) was dissolved in methanol (10 mL).Sodium acetate (0.30 g, 3.70 mmol) was added followed by 4 Å powderedmolecular sieves (0.40 g) and 4-fluoro-benzaldehyde (0.20 mL, 1.85mmol). Sodium cyanoborohydride (0.23 g, 3.70 mmol) was added and themixture was stirred at 25° C. for 16 h. The mixture was poured into amixture of saturated aqueous sodium bicarbonate solution (20 mL) andethyl acetate (30 mL). After shaking, both layers were passed through aplug of Celite. The organic layer was further washed with saturatedaqueous sodium bicarbonate solution (10 mL), saturated aqueous brinesolution (10 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product, rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylateas a clear oil. LC-MS (ESI) calcd for C₁₇H₂₂FNO₂ 291.36, found 292.2[M+H⁺].

f)rac-N-{3-[(1R,2R,7S,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]tridec-5-en-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide

rac-Methyl(1R,2R,4S,5S,6S,7R)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate(0.15 g, 0.49 mmol) was dissolved in anhydrous N,N-dimethylformamide (8mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.16 g, 0.49 mmol) was addedfollowed by N-methylmorpholine (0.11 mL, 1.03 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.10 g,0.51 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.20 mL, 1.47 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (50 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×50 mL), saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 20% ethyl acetate in dichloromethane) afforded thedesired product,rac-N-{3-[(1R,2R,7S,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]tridec-5-en-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide(0.23 g, 0.40 mmol, 82%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 0.44-1.28 (10H, m), 2.29 (1H, bs), 3.06 (3H, s), 3.82 (1H, d, J=11.6Hz), 4.40 (1H, d, J=14.0 Hz), 4.98 (1H, d, J=15.6 Hz), 7.12-7.60 (7H,m), 10.19 (1H, s). LC-MS (ESI) calcd for C₂₇H₂₇FN₄O₆S₂ 586.65, found587.5 [M+H⁺].

Example 52rac-N-{3-[(1R,2S,7R,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]trideca-5,12-dien-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide

a)rac-(1S,2S,4R,5R,6S,7R)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid

rac-(1R,2S,6R,7S,8S,10R)-4-Oxatetracyclo[5.3.2.0^(2,6).0^(8,10)]dodec-11-ene-3,5-dione(prepared as described in Example 51a, 0.86 g, 4.50 mmol) was dissolvedin methanol (20 mL). The reaction was stirred at 25° C. for 72 h. Themixture was concentrated to afford a clear oil. Purification by flashcolumn chromatography (Teledyne Isco RediSep column; 0 to 60% ethylacetate in hexanes) afforded the desired product,rac-(1S,2S,4R,5R,6S,7R)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid (0.94 g, 4.23 mmol, 94%) as a white solid. ¹H NMR (400 MHz, CDCl₃)δ: 0.06-0.17 (2H, m), 1.26-1.38 (2H, m), 2.74-2.85 (2H, m), 3.15-3.19(2H, m), 3.67 (3H, s), 5.85-5.86 (2H, m). LC-MS (ESI) calcd for C₁₂H₁₄O₄222.24, found 223.5 [M+H⁺].

b) rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate

rac-(1S,2S,4R,5R,6S,7R)-7-(Methoxycarbonyl)tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylicacid (0.68 g, 3.07 mmol) was dissolved in anhydrous tetrahydrofuran (20mL). The flask was degassed and backfilled with nitrogen and the mixturewas cooled to 0° C. Triethylamine (1.28 mL, 9.21 mmol) was addedfollowed by the dropwise addition of ethyl chloroformate (0.58 mL, 6.14mmol) with vigorous stirring. The mixture was stirred at 0° C. for 1 h.Sodium azide (0.60 g, 9.21 mmol) was dissolved in water (5 mL) and addedto the reaction mixture at 0° C. The mixture was stirred at 0° C. for 5min. The ice bath was removed. The mixture was warmed to 25° C. and wasstirred for 2 h. The mixture was poured into water (70 mL) and theproduct extracted into ethyl acetate (70 mL). The organic layer wasfurther washed with half-saturated aqueous sodium bicarbonate solution(2×30 mL), saturated aqueous brine solution (30 mL), dried overmagnesium sulfate, filtered, and concentrated in vacuo to afford a clearoil. The oil was dissolved in anhydrous benzene (10 mL) and refluxedwhile stirring under nitrogen for 2 h. Upon cooling to 25° C. thesolution was concentrated in vacuo to afford a slightly yellow oil. Theoil was dissolved in dichloromethane (10 mL) and benzyl alcohol (0.35mL, 3.38 mmol) was added followed by triethylamine (0.86 mL, 6.14 mmol).The mixture was refluxed under nitrogen for 16 h. Upon cooling to 25° C.the solution was concentrated in vacuo to afford a golden oil.Purification by flash column chromatography (Teledyne Isco RediSepcolumn; 0 to 20% ethyl acetate in hexanes) afforded the desired product,rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate(0.36 g, 1.10 mmol, 36%) as a clear oil. ¹H NMR (400 MHz, CDCl₃) δ:0.11-0.19 (2H, m), 0.13-1.34 (2H, m), 2.77-3.03 (3H, m), 3.62 (3H, s),4.01-4.07 (1H, m), 5.07-5.14 (2H, m), 5.79-5.85 (2H, m), 7.30-7.38 (5H,m). LC-MS (ESI) calcd for C₁₉H₂₁NO₄ 327.37, found 328.3 [M+H⁺].

c) rac-Methyl(1R,2R,4S,5S,6R,7S)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate

rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(benzyloxy)carbonyl]amino}tricyclo[3.2.2.0^(2,4)]non-8-ene-6-carboxylate(0.35 g, 1.07 mmol) was dissolved in ethyl acetate (10 mL). 5% Palladiumon carbon (0.04 g) was added. The flask was degassed and backfilled withhydrogen gas via balloon. The mixture was stirred at 25° C. for 16 h.The mixture was passed through a plug of Celite and the filtrate wasconcentrated in vacuo to afford the crude product, rac-Methyl(1R,2R,4S,5S,6R,7S)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate,as a thick clear oil, which was used in the next step without furtherpurification. LC-MS (ESI) calcd for C₁₁H₁₇NO₂ (free amine) 195.26, found196.3 [M+H⁺].

d) rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate

The crude rac-Methyl(1R,2R,4S,5S,6R,7S)-7-aminotricyclo[3.2.2.0^(2,4)]nonane-6-carboxylatewas dissolved in methanol (10 mL). Acetic acid (0.12 mL, 2.14 mmol) wasadded followed by 4-fluoro-benzaldehyde (0.12 mL, 1.07 mmol). Sodiumcyanoborohydride (0.14 g, 2.14 mmol) was added and the mixture wasstirred at 60° C. for 16 h. The mixture was cooled to 25° C. and pouredinto a mixture of saturated aqueous sodium bicarbonate solution (20 mL)and ethyl acetate (30 mL). After shaking, both layers were passedthrough a plug of Celite. The organic layer was further washed withsaturated aqueous sodium bicarbonate solution (10 mL), saturated aqueousbrine solution (10 mL), dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford the crude product, rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylate,as a clear oil, which was used in the next step without furtherpurification. LC-MS (ESI) calcd for C₁₇H₂₂FNO₂ 291.36, found 292.0[M+H⁺].

e)rac-N-{3-[(1R,2S,7R,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]trideca-5-en-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide

The crude rac-Methyl(1R,2R,4S,5S,6R,7S)-7-{[(4-fluorophenyl)methyl]amino}tricyclo[3.2.2.0^(2,4)]nonane-6-carboxylatewas dissolved in anhydrous N,N-dimethylformamide (8 mL).(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.22 g, 0.66 mmol) was addedfollowed by N-methylmorpholine (0.15 mL, 1.39 mmol). The mixture wasstirred until everything dissolved, approximately 5 min.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.13 g,0.69 mmol) was added and the mixture was stirred at 25° C. for 4 h.Triethylamine (0.28 mL, 1.98 mmol) was added and the mixture was stirredat 50° C. for 16 h. Upon cooling to 25° C., the solution was dilutedwith ethyl acetate (50 mL) and washed with 1.0M aqueous hydrochloricacid solution (2×50 mL), saturated aqueous brine solution (20 mL), driedover magnesium sulfate, filtered, and concentrated in vacuo to afford agolden oil. Purification by flash column chromatography (Teledyne IscoRediSep column; 0 to 20% ethyl acetate in dichloromethane) afforded thedesired product,rac-N-{3-[(1R,2S,7R,8S,9S,11R)-3-[(4-Fluorophenyl)methyl]-6-hydroxy-4-oxo-3-azatetracyclo[6.3.2.0^(2,7).0^(9,11)]trideca-5-en-5-yl]-1,1-dioxo-4H-1λ⁶,2,4-benzothiadiazin-7-yl}methanesulfonamide(0.50 g, 0.86 mmol, 80% over three steps), as a white powder. ¹H NMR(400 MHz, DMSO-d₆) δ: 0.33-1.34 (10H, m), 2.30 (1H, bs), 3.06 (3H, s),3.81 (1H, d, J=11.2 Hz), 4.48 (1H, d, J=15.6 Hz), 4.98 (1H, d, J=14.8Hz), 7.13-7.60 (7H, m), 10.19 (1H, s). LC-MS (ESI) calcd forC₂₇H₂₇FN₄O₆S₂ 586.65, found 587.3 [M+H⁺].

Example 53(1R,2S,7R,8S)-5-(1,1-Dioxo-7-pyrrolidin-1-yl-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-6-hydroxy-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

Pyrrolidine (15 μL, 0.180 mmol), sarcosine (N-methyl glycine) (3.2 mg,0.018 mmol), copper (I) iodide (3.4 mg, 0.018 mmol), and potassiumphosphate (57.3 mg, 0.270 mmol) were placed in a flask under a nitrogenatmosphere.(1R,2S,7R,8S)-6-Hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 34, 50 mg, 0.090 mmol) was dissolvedin N,N-dimethylformamide (1 mL) and added to the above mixture. Themixture was stirred at 80° C. for 26 h. Additional pyrrolidine (50 μL,0.60 mmol), sarcosine (N-methyl glycine) (10 mg, 0.056 mmol), and copper(I) iodide (15 mg, 0.079 mmol) were added and the mixture was stirred at80° C. for another 16 h until LC-MS analysis indicated completion of thereaction. The reaction was repeated at the same scale as described aboveand after stirring at 80° C. for 24 h, additional pyrrolidine (50 μL,0.60 mmol), sarcosine (N-methyl glycine) (10 mg, 0.056 mmol), and copper(I) iodide (15 mg, 0.079 mmol) were added and the mixture was stirred at80° C. for another 18 h until LC-MS analysis indicated completion of thereaction. Both batches were combined and filtered through a plug ofCelite. The filter cake was washed with 10% methanol in dichloromethane(2×2 mL) and N,N-dimethylformamide (2 mL). The filtrate was concentratedin vacuo and the residue was purified by prep-HPLC [Column Luna 5μ C18(2) 100 Å AXIA 150×21.2 mm, 5 micron, 30%-95% in 7 min @30 mL/min flowrate, 0.05% trifluoroacetic acid in acetonitrile/0.05% trifluoroaceticacid in water] to afford the desired product,(1R,2S,7R,8S)-5-(1,1-dioxo-7-pyrrolidin-1-yl-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-6-hydroxy-3-(3-methyl-butyl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(36.7 mg, 0.074 mmol, 40.9%), as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.92 (6H, d, J=5.5 Hz), 1.21-1.60 (9H, m), 1.96-1.99 (4H,m), 2.52-2.52 (1H, m), 2.62-2.65 (1H, m), 2.94-2.98 (1H, m), 3.03-3.10(1H, m), 3.27-3.30 (4H, m), 3.60-3.71 (2H, m), 6.71 (1H, d, J=2.3 Hz),6.90 (1H, dd, J₁=9.2 Hz, J₂=2.7 Hz), 7.41 (1H, d, J=9.5 Hz). LC-MS (ESI)calcd for C₂₆H₃₄N₄O₄S 498.23, found 499.4 [M+H⁺]. Anal. calcd forC₂₆H₃₄N₄O₄S.0.5 H₂O: C, 61.51; H, 6.95; N, 11.04; found: C, 61.45, H,6.74; N, 10.91.

Example 54 Pyridine-3-sulfonic acid{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide

(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-5-(7-iodo-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 19, 100 mg, 0.17 mmol),pyridine-3-sulfonic acid amide (81 mg, 0.51 mmol), sarcosine (N-methylglycine) (23 mg, 0.26 mmol), copper (I) iodide (33 mg, 0.17 mmol), andpotassium phosphate (216 mg, 1.02 mmol) were combined and dissolved inN,N-dimethylformamide (10 mL). The flask was degassed and backfilledwith nitrogen (3×). The reaction was stirred at 100° C. for 4 h. Themixture was cooled to 25° C., diluted with ethyl acetate (20 mL), andextracted with saturated aqueous sodium bicarbonate solution (2×20 mL).The combined organic layers were dried over magnesium sulfate, filtered,and concentrated in vacuo. Purification by flash column chromatography(Teledyne Isco RediSep column; 0 to 8% methanol in dichloromethane)afforded the desired product, pyridine-3-sulfonic acid{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-amide(56 mg, 0.09 mmol, 54%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆) δ:1.38-1.58 (6H, m), 2.61 (1H, s), 3.00 (1H, d, J=7.6 Hz), 3.52 (1H, d,J=8.4 Hz), 4.40 (1H, d, J=15.6 Hz), 4.94 (1H, d, J=15.2 Hz), 7.13 (2H,t, J=8.4 Hz), 7.29-7.32 (2H, m), 7.42-7.50 (3H, m), 7.60-7.63 (1H, m),8.11 (1H, d, J=7.6 Hz), 8.79 (1H, d, J=4.4 Hz), 8.87 (1H, s), 10.94 (1H,s), 13.99 (1H, s). LC-MS (ESI) calcd for C₂₉H₂₆FN₅O₆S₂ 623.68, found624.2 [M+H⁺].

Example 55(1R,2S,7R,8S)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamide

a) Benzyl[N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamoyl]carbamate

Benzyl alcohol (35 μL, 0.338 mmol) was dissolved in dichloromethane (3mL) and cooled to 0° C. Chlorosulfonyl isocyanate (29.4 μL, 0.338 mmol)was added and the mixture was stirred at 0° C. for 2.5 h. Triethylamine(47 μL, 0.34 mmol) followed by a solution of(1R,2S,7R,8S)-5-(7-amino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 30, 81.4 mg, 0.169 mmol) indichloromethane (2 mL) were added at 0° C. and the mixture was stirredat 25° C. for 17 h. The mixture was extracted with water (2×2 mL) andsaturated aqueous brine solution (2 mL). The organic layer wasconcentrated in vacuo and further dried for 16 h under high vacuum toafford the crude product, benzyl[N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamoyl]carbamate,which was used in the next step without any further purification. LC-MS(ESI) calcd for C₃₂H₃₀FN₅O₈S₂ 695.15, found 696.6 [M+H⁺].

b)(1R,2S,7R,8S)—N-{3-[3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamide

The crude benzyl[N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamoyl]carbamatewas dissolved in methanol (5 mL) and the mixture was degassed andbackfilled with nitrogen (3×). Palladium on carbon (10% dry, 180 mg) wasadded and the mixture was degassed and backfilled with hydrogen gas viaballoon. The mixture was stirred at 25° C. for 16 h. The mixture waspassed through a plug of Celite and the filtrate was concentrated invacuo to afford the crude product, which was further purified byprep-HPLC [Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron,30%-95% in 7 min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct,(1R,2S,7R,8S)—N-{3-[3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-sulfamide(55.7 mg, 0.099 mmol, 58.7% over two steps), as a white solid. ¹H NMR(400 MHz, DMSO-d₆) δ: 1.19-1.24 (2H, m), 1.40-1.60 (4H, m), 2.51-2.54(1H, m), 2.64-2.65 (1H, m), 3.04 (1H, d, J=9.3 Hz), 3.53 (1H, d, J=9.3Hz), 4.42 (1H, d, J=15.7 Hz), 4.96 (1H, d, J=15.6 Hz), 7.15 (2H, m),7.31-7.34 (H, m), 7.43 (1H, dd, J₁=9.4 Hz, J₂=2.3 Hz), 7.51-7.55 (2H,m), 9.96 (1H, s). LC-MS (ESI) calcd for C₂₄H₂₄FN₅O₆S 561.12, found 562.5[M+H⁺]. Anal. calcd for C₂₄H₂₄FN₅O₆S.0.5 H₂O: C, 50.51; H, 4.42; N,12.27; found: C, 50.42, H, 4.35; N, 11.90.

Example 56(1R,2S,7R,8S)—N-[3-(3-Benzyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

a) (1S,2R,3S,4R)-3-Benzylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester

Benzaldehyde (0.454 mL, 4.47 mmol) and 10 drops glacial acetic acid wereadded sequentially to a solution of(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 0.82 g, 4.47 mmol) inmethanol (15 mL) at 25° C. Sodium cyanoborohydride (0.703 g, 11.2 mmol)was added, and the reaction mixture was stirred at 25° C. for 2 h, andthen was partitioned between half-saturated aqueous sodium bicarbonatesolution (150 mL) and ethyl acetate (2×150 mL). The organic layers weredried over sodium sulfate, filtered and concentrated in vacuo. Theresidue was purified by flash column chromatography (Teledyne IscoRediSep column; 0 to 35% ethyl acetate in hexanes) to afford the desiredproduct, (1S,2R,3S,4R)-3-benzylamino-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.875 g, 3.22 mmol, 72%), as a clear oil. ¹H NMR (400MHz, CDCl₃) δ: 1.06-1.16 (2H, m), 1.20-1.23 (2H, m), 1.28 (3H, t, J=7.0Hz), 1.46-1.61 (2H, m), 1.93-1.97 (1H, m), 2.33-2.34 (1H, m), 2.43-2.44(1H, m), 2.59-2.62 (1H, m), 2.98-3.00 (1H, m), 3.71 (1H, d, J=14.0 Hz),3.85 (1H, d, J=13.3 Hz), 4.14 (2H, q, J=7.3 Hz), 7.20-7.24 (1H, m),7.27-7.33 (4H, m).

b)(1R,2S,7R,8S)—N-[3-(3-Benzyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.200 g, 0.600 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.121 g,0.631 mmol) and N-methylmorpholine (0.139 mL, 1.26 mmol) were addedsequentially to a solution of(1S,2R,3S,4R)-3-benzylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester (0.164 g, 0.600 mmol) in N,N-dimethylformamide (5 mL) at 25°C. The reaction mixture was stirred at 25° C. for 2 h, and then waspartitioned between 1.0M aqueous hydrochloric acid solution (100 mL) andethyl acetate (2×100 mL). The organic layers were dried over sodiumsulfate, filtered and concentrated in vacuo. The residue was dissolvedin ethanol (15 mL) at 25° C. A 21 wt. % solution of sodium ethoxide inethanol (0.972 mL, 3.00 mmol) was added and the reaction mixture washeated at 60° C. for 1 h. After cooling to 25° C., the reaction mixturewas partitioned between a 1.0M aqueous hydrochloric acid solution (100mL) and ethyl acetate (2×100 mL). The organic layers were dried oversodium sulfate, filtered and concentrated in vacuo. The residue waspurified by flash column chromatography (Teledyne Isco RediSep column;40 to 100% ethyl acetate in hexanes) to afford the desired product,(1R,2S,7R,8S)—N-[3-(3-benzyl-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(0.120 g, 0.222 mmol, 37%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.15-1.21 (2H, m), 1.39-1.61 (4H, m), 2.52 (1H, bs), 2.64 (1H, bs),3.05 (3H, s), 3.30 (2H, bs), 3.54 (1H, d, J=9.5 Hz), 4.43 (1H, d, J=16.4Hz), 4.99 (1H, d, J=15.5 Hz), 7.23-7.28 (2H, m), 7.33 (2H, m), 7.48-7.51(2H, m), 7.55-7.57 (2H, m), 10.17 (1H, s). LC-MS (ESI) calcd forC₂₅H₂₆N₄O₆S₂ 542.13, found 543.2 [M+H⁺].

Example 57(1R,2S,7R,8S)—N-[3-(6-Hydroxy-3-isobutyl-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

a) (1S,2R,3S,4R)-3-Isobutylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester

Isovaleraldehyde (0.374 mL, 4.10 mmol) and 10 drops glacial acetic acidwere added sequentially to a solution of(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 0.750 g, 4.09 mmol) inmethanol (12 mL) at 25° C. Sodium cyanoborohydride (0.643 g, 10.2 mmol)was added, and the reaction mixture was stirred at 25° C. for 2 h, andthen was partitioned between half-saturated aqueous sodium bicarbonatesolution (150 mL) and ethyl acetate (2×150 mL). The organic layers weredried over sodium sulfate, filtered and concentrated in vacuo to affordthe desired product,(1S,2R,3S,4R)-3-isobutylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester, as a clear oil, which was used in the next step without anyfurther purification.

b)(1R,2S,7R,8S)—N-[3-(6-Hydroxy-3-isobutyl-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.317 g, 0.951 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.192 g,1.00 mmol) and N-methylmorpholine (0.220 mL, 2.00 mmol) were addedsequentially to a solution of crude(1S,2R,3S,4R)-3-isobutylamino-bicyclo[2.2.1]heptane-2-carboxylic acidethyl ester (0.228 g, 0.952 mmol) in N,N-dimethylformamide (4 mL) at 25°C. The reaction mixture was stirred at 25° C. for 2 h, and then waspartitioned between 1.0M aqueous hydrochloric acid solution (100 mL) andethyl acetate (2×100 mL). The organic layers were dried over sodiumsulfate, filtered and concentrated in vacuo. The residue was dissolvedin ethanol (15 mL) at 25° C. A 21 wt. % solution of sodium ethoxide inethanol (1.23 mL, 3.80 mmol) was added and the reaction mixture washeated at 60° C. for 1.5 h. After cooling to 25° C., the reactionmixture was partitioned between 1.0M aqueous hydrochloric acid solution(150 mL) and ethyl acetate (2×150 mL). The organic layers were driedover sodium sulfate, filtered and concentrated in vacuo. The residue waspurified by flash column chromatography (Teledyne Isco RediSep column;40 to 100% ethyl acetate in hexanes) to afford the desired product,(1R,2S,7R,8S)—N-[3-(6-hydroxy-3-isobutyl-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl)-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide(0.096 g, 0.190 mmol, 20%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 0.83 (3H, d, J=7.2 Hz), 0.93 (3H, d, J=7.1 Hz), 1.19-1.21 (1H, m),1.28-1.32 (1H, m), 1.43-1.63 (3H, m), 2.09-2.15 (1H, m), 2.54 (1H, bs),2.61-2.61 (1H, m), 2.74-2.79 (1H, m), 3.05 (3H, s), 3.30 (1H, bs), 3.60(1H, d, J=9.2 Hz), 3.73-3.79 (1H, m), 7.49-7.52 (1H, m), 7.56-7.58 (2H,m), 10.17 (1H, s). LC-MS (ESI) calcd for C₂₂H₂₈N₄O₆S₂ 508.15, found509.4 [M+H⁺].

Example 58(1R,2S,7R,8S)—N-{3-[3-(3-Chloro-4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(3-Chloro-4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

3-Chloro-4-fluoro-benzaldehyde (0.710 mL, 4.48 mmol) and 10 dropsglacial acetic acid were added sequentially to a solution of(1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 0.82 g, 4.47 mmol) inmethanol (15 mL) at 25° C. Sodium cyanoborohydride (0.709 g, 11.3 mmol)was added, and the reaction mixture was stirred at 25° C. for 2 h, andthen was partitioned between half-saturated aqueous sodium bicarbonatesolution (150 mL) and ethyl acetate (2×150 mL). The organic layers weredried over sodium sulfate, filtered and concentrated in vacuo. Theresidue was purified by flash column chromatography (Teledyne IscoRediSep column; 0 to 35% ethyl acetate in hexanes) to afford the desiredproduct,(1S,2R,3S,4R)-3-(3-chloro-4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (1.12 g, 3.44 mmol, 77%), as a clear oil. ¹H NMR (400MHz, CDCl₃) δ: 1.04-1.16 (2H, m), 1.18-1.24 (2H, m), 1.29 (3H, t, J=7.4Hz), 1.44-1.61 (2H, m), 1.91-1.94 (1H, m), 2.27-2.28 (1H, m), 2.42-2.43(1H, m), 2.60 (1H, d, J=7.7 Hz), 2.91 (1H, d, J=8.8 Hz), 3.64 (1H, d,J=13.9 Hz), 3.79 (1H, d, J=14.2 Hz), 4.14 (2H, q, J=7.0 Hz), 7.02-7.06(1H, m), 7.13-7.17 (1H, m), 7.36-7.38 (1H, m).

b)(1R,2S,7R,8S)—N-{3-[3-(3-Chloro-4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

(7-Methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 0.258 g, 0.774 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.156 g,0.814 mmol) and N-methylmorpholine (0.170 mL, 1.55 mmol) were addedsequentially to a solution of(1S,2R,3S,4R)-3-(3-chloro-4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.252 g, 0.773 mmol) in N,N-dimethylformamide (4 mL)at 25° C. The reaction mixture was stirred at 25° C. for 2 h, and thenwas partitioned between 1.0M aqueous hydrochloric acid solution (100 mL)and ethyl acetate (2×100 mL). The organic layers were dried over sodiumsulfate, filtered and concentrated in vacuo. The residue was dissolvedin ethanol (20 mL) at 25° C. A 21 wt. % solution of sodium ethoxide inethanol (1.00 mL, 3.09 mmol) was added and the reaction mixture washeated at 60° C. for 1 h. After cooling to 25° C., the reaction mixturewas partitioned between 1.0M aqueous hydrochloric acid solution (100 mL)and ethyl acetate (2×100 mL). The organic layers were dried over sodiumsulfate, filtered and concentrated in vacuo. The residue was purified byflash column chromatography (Teledyne Isco RediSep column; 40 to 100%ethyl acetate in hexanes) to afford the desired product,(1R,2S,7R,8S)—N-{3-[3-(3-chloro-4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.196 g, 0.332 mmol, 43%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ: 1.04-1.21 (4H, m), 1.40-1.60 (3H, m), 2.54-2.63 (1H, m), 3.05 (3H,s), 3.31-3.40 (2H, m), 3.55-3.58 (1H, m), 4.45 (1H, d, J=16.1 Hz), 4.91(1H, d, J=14.7 Hz), 7.31-7.35 (2H, m), 7.51-7.57 (4H, m), 10.17 (1H, s),13.96 (1H, s). LC-MS (ESI) calcd for C₂₅H₂₄ClFN₄O₆S₂ 594.08, found 595.3[M+H⁺].

Example 59(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide

(rac-di-exo)-N-{3-[6-Hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 25, 67 mg, 0.128 mmol) was dissolvedin N,N-dimethylformamide (2 mL). Potassium carbonate (35 mg, 0.25 mmol)and iodomethane (0.008 mL, 0.128 mmol) were added sequentially. Thereaction was stirred at 25° C. for 2 h. The reaction was quenched viathe addition of 1.0 M aqueous hydrochloric acid solution (20 mL). Themixture was extracted with ethyl acetate (2×100 mL). The combinedorganic layers were washed with saturated aqueous brine solution (50mL), dried over sodium sulfate, filtered, and concentrated in vacuo toafford an oil. Purification by flash column chromatography (TeledyneIsco RediSep column; 0 to 100% ethyl acetate in hexanes) afforded thedesired product,(rac-di-exo)-N-{3-[6-hydroxy-3-(3-methyl-butyl)-4-oxo-11-oxa-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-N-methyl-methanesulfonamide(36 mg, 0.067 mmol, 52%), as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ:0.92 (6H, d, J=6.2 Hz), 1.16-1.65 (7H, m), 2.79-2.93 (2H, m), 2.96 (3H,s), 3.26 (3H, s), 3.62-3.69 (1H, m), 3.76-3.83 (1H, m), 4.60 (2H, d,J=21.8 Hz), 7.27 (1H, d, J=7.3 Hz), 7.55 (1H, d, J=8.5 Hz), 7.65 (1H,s). LC-MS (ESI) calcd for C₂₃H₃₀N₄O₇S₂ 538.16, found 539.4 [M+H⁺].

Example 60N-{3-[(1R,2S,7R,8S)-3-(3-Fluoro-4-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(3-Fluoro-4-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

(1S,2R,3S,4R)-3-Amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 300 mg, 1.637 mmol) wassuspended in ethanol (5 mL). 3-Fluoro-4-methyl-benzaldehyde (0.2 mL,1.637 mmol) was added followed by glacial acetic acid (0.1 mL, 3.724mmol) and 4 Å powdered molecular sieves (0.6 g). Sodium cyanoborohydride(0.593 g, 9.406 mmol) was added and the mixture was stirred undernitrogen at 50° C. for 18 h. Upon cooling, the mixture was filteredthrough Celite. The filtrate was diluted with half-saturated aqueoussodium bicarbonate solution (50 mL) and extracted with ethyl acetate(2×75 mL). The layers were separated and the organic layer was washedwith saturated aqueous brine solution, dried over sodium sulfate,filtered, and concentrated in vacuo. Further purification by flashcolumn chromatography (Teledyne Isco RediSep column; 1^(st) column: 0 to25% ethyl acetate in hexanes; 2^(nd) column: 0 to 20% ethyl acetate inhexanes) afforded the desired product,(1S,2R,3S,4R)-3-(3-fluoro-4-methyl-benzylamino)-bicyc[2.2.1]heptane-2-carboxylic acid ethyl ester (221 mg, 0.724 mmol, 44%),as a clear thick oil. ¹H NMR (400 MHz, CDCl₃) δ: 1.04-1.15 (2H, m), 1.21(1H, d, J=10.1 Hz), 1.29 (3H, t, J=7.1 Hz), 1.42-1.63 (4H, m), 1.94 (1H,dt, J₁=10.2 Hz, J₂=2.0 Hz), 2.25 (3H, s), 2.43 (1H, d, J=3.5 Hz), 2.59(1H, dd, J₁=8.6 Hz, J₂=1.6 Hz), 2.93 (1H, dd, J₁=8.6 Hz, J₂=1.6 Hz),3.65 (1H, d, J=13.4 Hz), 3.79 (1H, d, J=14.2 Hz), 4.15 (2H, q, J=7.1Hz), 6.94 (1H, d, J=7.9 Hz), 6.98 (1H, d, J=10.9 Hz), 7.08 (1H, t, J=7.8Hz). LC-MS (ESI) calcd for C₁₈H₂₄FNO₂ 305.18, found 305.9 [M+H⁺].

b)(1S,2R,3S,4R)-3-{(3-Fluoro-4-methyl-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

To a solution of(1S,2R,3S,4R)-3-(3-fluoro-4-methyl-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (92 mg, 0.30 mmol),(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 100 mg, 0.30 mmol) and1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (60 mg,0.315 mmol) in N,N-dimethylformamide (4.0 mL) was addedN-methylmorpholine (0.07 mL, 0.63 mmol). After stirring at 25° C. for 3h, the mixture was poured into a 1.0M aqueous hydrochloric acid solutionand extracted with ethyl acetate (2×100 mL). The combined organic layerswere washed with saturated aqueous brine solution, dried over sodiumsulfate, filtered, and concentrated in vacuo to afford the crudeproduct,(1S,2R,3S,4R)-3-{(3-fluoro-4-methyl-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as an orange oil. The crude product was used in thenext step without further purification. LC-MS (ESI) calcd forC₂₈H₃₃FN₄O₇S₂ 620.18, found 621.4 [M+H⁺].

c)N-{3-[(1R,2S,7R,8S)-3-(3-Fluoro-4-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

The crude(1S,2R,3S,4R)-3-{(3-fluoro-4-methyl-benzyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester was dissolved in ethanol (5 mL) at 25° C. A 21 wt. %solution of sodium ethoxide in ethanol (0.224 mL, 0.60 mmol) was addedand the reaction mixture was heated at 60° C. for 90 min. After coolingto 25° C., the reaction mixture stirred for 18 h at 25° C. The mixturewas partitioned between a 1.0M aqueous hydrochloric acid solution (100mL) and ethyl acetate (2×100 mL). The organic layers were washed withsaturated aqueous brine solution, dried over sodium sulfate, filteredand concentrated in vacuo. The residue was purified by flash columnchromatography (Teledyne Isco RediSep column; 1^(st) column: 0 to 10%ethyl acetate in hexanes; 2^(nd) column: 30 to 80% ethyl acetate inhexanes) to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(3-fluoro-4-methyl-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(53 mg, 0.092 mmol, 31% over two steps), as a white solid. ¹H NMR (400MHz, DMSO-d₆) δ: 1.12-1.21 (2H, m), 1.39-1.64 (4H, m), 2.20 (3H, s),2.63 (1H, bs), 3.02 (1H, d, J=9.9 Hz), 3.06 (3H, s), 3.54 (1H, d, J=9.3Hz), 4.42 (1H, d, J=15.5 Hz), 4.92 (1H, d, J=15.5 Hz), 7.01-7.07 (2H,m), 7.23 (1H, t, J=8.1 Hz), 7.49-7.58 (3H, m), 10.17 (1H, s). LC-MS(ESI) calcd for C₂₆H₂₇FN₄O₆S₂ 574.14, found 575.4 [M+H⁺].

Example 61(1R,2S,7R,8S)-5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a) 2-Amino-5-bromo-pyridine-3-sulfonic acid amide

5-Bromo-pyridin-2-ylamine (10 g, 57.8 mmol) was added to chlorosulfonicacid (40 mL, 602 mmol) while stirring at 25° C. The mixture was heatedat 160° C. while stirring for 3 h. Upon cooling to 25° C., the brownsolution was carefully poured over ice (˜500 g). The resultingprecipitate was collected by vacuum filtration and rinsed with water toafford the intermediate, 2-amino-5-bromo-pyridine-3-sulfonyl chloride,as a beige solid. The solid was suspended in a 15% aqueous ammoniumhydroxide solution. The reaction mixture stirred for 45 min. Everythingwas completely dissolved at this point. The mixture was cooled to 0° C.and the pH was adjusted to ˜8 by the careful addition of 12.0M aqueoushydrochloric acid solution. A solid precipitated and was collected byvacuum filtration, rinsed with water (2×50 mL) and dried in vacuo toafford the desired product, 2-amino-5-bromo-pyridine-3-sulfonic acidamide (7.48 g, 29.7 mmol, 51%), as a light beige solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 6.69 (2H, bs), 7.54 (2H, bs), 7.91 (1H, d, J=2.5 Hz), 8.22(1H, d, J=2.3 Hz).

b) N-(5-Bromo-3-sulfamoyl-pyridin-2-yl)-malonamic acid ethyl ester

2-Amino-5-bromo-pyridine-3-sulfonic acid amide (2.5 g, 9.9 mmol) wasdissolved in 1,4-dioxane (50 mL). Chlorocarbonyl-acetic acid ethyl ester(1.9 mL, 14.85 mmol) was added and the mixture was heated at 90° C.while stirring for 2 h. Upon cooling to 25° C., the mixture was pouredinto half-saturated aqueous sodium bicarbonate solution (250 mL). Asolid precipitated and was collected by vacuum filtration, rinsed withwater (50 mL) and dried in vacuo to afford the desired product,N-(5-bromo-3-sulfamoyl-pyridin-2-yl)-malonamic acid ethyl ester (2.5 g,6.82 mmol, 69%), as a light beige solid. ¹H NMR (400 MHz, DMSO-d₆) δ:1.19 (3H, t, J=7.0 Hz), 3.67 (2H, s), 4.10 (2H, q, J=7.3 Hz), 7.78 (2H,bs), 8.33 (1H, d, J=2.3 Hz), 8.70 (1H, d, J=2.4 Hz), 9.85 (1H, bs).

c)(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-aceticacid ethyl ester

In a sealed tube, N-(5-Bromo-3-sulfamoyl-pyridin-2-yl)-malonamic acidethyl ester (2.2 g, 6.0 mmol) was suspended in toluene (44 mL) andtriethylamine (11 mL, 78.9 mmol) was added. The mixture was heated at110° C. while stirring for 30 min. The solid was completely dissolved,yet an immiscible, oily residue was observed along the bottom of theflask. Upon cooling to 25° C., ethyl acetate (˜50 mL) was added.Everything became miscible. The solution was concentrated in vacuo toafford a golden oil. The oil was dissolved in methanol (˜50 mL) andconcentrated in vacuo to afford the crude product,(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-aceticacid ethyl ester (2.18 g, >6.0 mmol, 100%, still contained somesolvent), as a golden oil. LC-MS (ESI) calcd for C₁₀H₁₀BrN₃O₄S 346.96,found 348.1 (100%), 349.2 (10%), 350.2 (99%) [M+H⁺].

d) Sodium-(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-acetate

The crude(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-aceticacid ethyl ester (1.14 g, 3.27 mmol) was dissolved in methanol (20 mL).Solid sodium hydroxide (0.392 g, 9.8 mmol) was added followed by water(10 mL). Everything was completely dissolved within ˜5 minutes. After˜20 minutes, a solid began to precipitate. The mixture continued to stirfor 10 min. The product was collected by vacuum filtration, rinsed withmethanol (˜5 mL) and dried under vacuum to afford the desired product,sodium-(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-acetate(0.48 g, 1.5 mmol, 46%), as a white solid. ¹H NMR (400 MHz, D₂O) δ: 3.30(2H, s), 8.31 (1H, d, J=2.2 Hz), 8.57 (1H, d, J=1.6 Hz). Note:Successive NMR acquisitions of the product in DMSO-d₆ indicateddecarboxylation over a period of ˜30 min to afford7-bromo-3-methyl-4H-pyrido[2,3-e][1,2,4]thiadiazine 1,1-dioxide.However, the product appears to be stable as the solid sodium salt form.

e)(1R,2S,7R,8S)-5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

(1S,2R,3S,4R)-3-(4-Fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 6l, 0.213 g, 0.73mmol),sodium-(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-acetate(0.25 g, 0.73 mmol) andO-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (0.305 g, 0.803 mmol) were combined and dissolved inN,N-dimethylformamide (1.5 mL). The mixture was shaken vigorously forabout 1 min until everything dissolved. The solution continued to stirfor 1 h. Triethylamine (0.5 mL, 3.65 mmol) was added and the mixture wasstirred at 50° C. for 16 h. The mixture was concentrated in vacuo to avolume of ˜5 mL. Water (˜20 mL) was added and the product precipitated.The solid was collected by vacuum filtration and purified by flashcolumn chromatography (Merck silical gel 60, 40-63 μm; 35% ethyl acetatein hexanes) to afford the desired product, (0.24 g, 0.44 mmol, 60%) as awhite, brittle foam. A portion of the product (0.07 g, 0.128 mmol) wasrecrystallized from n-propanol (0.5 mL) to afford the pure desiredproduct,5-(7-bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.044 g, 0.081 mmol, 63%), as a yellow powder. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.13-1.23 (2H, m), 1.38-1.60 (4H, m), 2.51 (1H, s), 2.63(1H, s), 3.00 (1H, d, J=8.4 Hz), 3.55 (1H, d, J=9.4 Hz), 4.41 (1H, d,J=14.7 Hz), 4.95 (1H, d, J=15.4 Hz), 7.14 (2H, t, J=9.0 Hz), 7.33 (2H,dd, J₁=8.6 Hz, J₂=5.5 Hz), 8.66 (1H, s), 8.82 (1H, s). LC-MS (ESI) calcdfor C₂₃H₂₀BrFN₄O₄S 546.04, found 547.2 [M+H⁺].

Example 62N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-7-yl}-methanesulfonamide

Methane sulfonamide (0.348 g, 3.66 mmol), potassium phosphate (0.078 g,0.366 mmol), L-proline (0.021 g, 0.183 mmol) and copper iodide (0.035 g,0.183 mmol) were combined and suspended in dimethylsulfoxide (0.5 mL).The flask was degassed and backfilled with argon. The mixture washeated, while stirring, at 110° C. for 5 min.5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 62, 0.1 g, 0.183 mmol) was dissolvedin dimethylsulfoxide (0.5 mL) and transferred to the reaction mixture.The mixture was stirred at 110° C. for 16 h. Upon cooling, the mixturewas poured into ethyl acetate (150 mL) and saturated aqueous ammoniumchloride solution (100 mL). The mixture was shaken and everything waspassed through a plug of Celite. The organic layer was separated fromthe filtrate, washed with saturated aqueous ammonium chloride solution(50 mL), saturated aqueous brine solution (50 mL), dried over magnesiumsulfate, filtered and concentrated in vacuo to a brown waxy solid. Flashcolumn chromatography (Merck silica gel 60, 40-63 μm; 0 to 50% ethylacetate in dichloromethane) afforded the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-7-yl}-methanesulfonamide(0.018 g, 0.032 mmol, 17.5%), as a beige solid. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.04-1.52 (6H, m), 2.37-2.41 (1H, m), 2.53-2.56 (1H, m),3.05 (3H, s), 3.25-3.34 (2H, m), 4.25 (1H, d, J=15.1 Hz), 4.93 (1H, d,J=15.0 Hz), 7.11 (2H, t, J=9.1 Hz), 7.27 (2H, dd, J₁=8.7 Hz, J₂=5.4 Hz),7.81 (1H, s), 8.36 (1H, s), 10.01 (1H, bs). LC-MS (ESI) calcd forC₂₄H₂₄FN₅O₆S₂ 561.12, found 562.4 [M+H⁺].

Example 63(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[2,3e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(prepared as described in Example 62, 0.07 g, 0.128 mmol) was dissolvedin methanol (8 mL). Ammonium formate (0.2 g, 3.17 mmol) followed by 10%palladium on carbon (wet, 0.1 g) were added. The mixture stirred at 50°C. for 3 h. The mixture was filtered through Celite and the filtrate wasconcentrated in vacuo. The residue was dissolved in water (5 mL) andethyl acetate (50 mL). The mixture was shaken and the layers wereseparated. The organic layer was dried over magnesium sulfate, filteredand concentrated in vacuo to a clear oil. Flash column chromatography(Merck silica gel 60, 40-63 μm; 20 to 40% ethyl acetate in hexanes)afforded the desired product,(1R,2S,7R,8S)-5-(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.0441 g, 0.094 mmol, 74%), as a white, brittle foam. ¹H NMR (400 MHz,DMSO-d₆) δ: 1.11-1.61 (6H, m), 2.51 (1H, s), 2.63 (1H, s), 2.99 (1H, d,J=9.4 Hz), 3.54 (1H, d, J=9.4 Hz), 4.41 (1H, d, J=15.7 Hz), 4.95 (1H, d,J=15.4 Hz), 7.14 (2H, t, J=8.6 Hz), 7.33 (2H, dd, J₁=8.6 Hz, J₂=5.4 Hz),7.51 (1H, dd, J₁=7.8 Hz, J₂=4.5 Hz), 8.34 (1H, d, J=7.6 Hz), 8.68 (1H,dd, J₁=4.6 Hz, J₂=1.7 Hz). LC-MS (ESI) calcd for C₂₃H₂₁FN₄O₄S 468.13,found 469.4 [M+H⁺].

Example 64(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a) 4-Azido-pyridine-3-sulfonic acid amide

4-Chloro-pyridine-3-sulfonic acid amide (4 g, 20.77 mmol) and sodiumazide (13.7 g, 210 mmol) were combined. Anhydrous N,N-dimethylformamide(80 mL) and water (30 mL) were added. The mixture was stirred at 90° C.for 2 h. Upon cooling, the mixture was diluted with saturated aqueousammonium chloride solution (200 mL). The product was extracted intoethyl acetate (6×200 mL). The combined organic layers were dried overmagnesium sulfate, filtered and concentrated in vacuo to afford thedesired product, 4-azido-pyridine-3-sulfonic acid amide (3.75 g, 18.83mmol, 91%), as a pale yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 7.54(2H, bs), 7.57 (1H, d, J=5.4 Hz), 8.68 (1H, d, J=5.5 Hz), 8.81 (1H, s).

b) 4-Amino-pyridine-3-sulfonic acid amide

4-Azido-pyridine-3-sulfonic acid amide (3.75 g, 18.83 mmol) wasdissolved in methanol (80 mL). Sodium borohydride (0.712 g, 18.83 mmol)was carefully added portionwise. Vigorous effervescence was observed.The mixture continued to stir at 25° C. for 25 min. The mixture wasconcentrated in vacuo to a thick yellow sludge. The residue wasdissolved in a mixture of ethyl acetate (200 mL) and saturated aqueousammonium chloride solution (200 mL). The aqueous layer was backextracted with ethyl acetate (6×200 mL). The combined organic layerswere dried over magnesium sulfate, filtered and concentrated in vacuo toafford the desired product, 4-amino-pyridine-3-sulfonic acid amide (1.8g, 10.4 mmol, 55%), as a pale yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ:6.63 (2H, bs), 6.68 (1H, d, J=5.4 Hz), 7.40 (2H, bs), 8.06 (1H, d, J=5.5Hz), 8.43 (1H, s).

c)(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-aceticacid methyl ester

4-amino-pyridine-3-sulfonic acid amide (0.25 g, 1.44 mmol) was suspendedin malonic acid dimethyl ester (5 mL, 43.75 mmol). The flask was purgedwith nitrogen and the mixture was stirred at 180° C. for 60 min. Uponcooling, the mixture was diluted with ethyl acetate (5 mL) causingimmediate precipitation of an undesired side product. The solid wasremoved by vacuum filtration and rinsed with ethyl acetate (2 mL). Thefiltrate was passed through a plug of silica gel, eluting with ethylacetate followed by 5% methanol in ethyl acetate. The fractionscontaining the 5% methanol in ethyl acetate were combined andconcentrated in vacuo to afford the desired product,(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-aceticacid methyl ester (0.135 g, 0.529 mmol, 37%) as a yellow oil. LC-MS(ESI) calculated for C₉H₉N₃O₄S 255.03, found 256.0 [M+H⁺].

d)Sodium-(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-acetate

(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-aceticacid methyl ester (0.13 g, 0.51 mmol) was dissolved in methanol (3 mL).Sodium hydroxide (0.08 g, 2.0 mmol) was dissolved in water (1.5 mL). Thesolutions were combined and were stirred at 25° C. for 3 h. The mixturewas concentrated in vacuo to afford the desired product,sodium-(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-acetate,as a yellow oil. LC-MS (ESI) calculated for C₈H₇N₃O₄S (free acid)241.02, found 242.2 [M+H⁺].

e)(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

The crudesodium-(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-acetate,(1S,2R,3S,4R)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (prepared as described in Example 6l, 0.149 g, 0.51mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N′-tetramethyluroniumhexafluorophosphate (0.194 g, 0.51 mmol) were combined and dissolved inanhydrous N,N-dimethylformamide (1 mL). The mixture was stirred at 25°C. for 1 h. Triethylamine (0.35 mL, 2.6 mmol) was added and the mixturewas stirred at 50° C. for 16 h. Upon cooling, the mixture was dilutedwith ethyl acetate (100 mL), washed with saturated aqueous ammoniumchloride solution (2×25 mL) and saturated aqueous brine solution (25mL). The organic layer was dried over magnesium sulfate, filtered andconcentrated in vacuo. Purification by flash column chromatography(Merck silica gel 60, 40-63 μm, 75% to 100% ethyl acetate in hexanes)followed trituration of the resulting oil with a 1:1 mixture of hexanesand diethyl ether (2 mL) afforded the desired product,(1R,2S,7R,8S)-5-(1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.0184 g, 0.039 mmol, 7.7% over two steps), as a pale yellow powder.LC-MS (ESI) calculated for C₂₃H₂₁FN₄O₄S 468.13, found 469.2 [M+H⁺]. ¹HNMR (400 MHz, DMSO-d₆) δ: 1.01-1.54 (6H, m), 2.35 (1H, d, J=3.7 Hz),2.46 (1H, s), 2.53 (1H, s), 3.26 (1H, d, J=9.5 Hz), 4.23 (1H, d, J=15.8Hz), 4.94 (1H, d, J=15.0 Hz), 7.08-7.13 (3H, m), 7.24-7.28 (2H, m), 8.44(1H, d, J=4.8 Hz), 8.67 (1H, s).

Example 65(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[3,2-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

a) 2-Benzylsulfanyl-3-nitro-pyridine

Ethanol (150 mL) was added to a mixture of 2-chloro-3-nitro-pyridine (5g, 31.54 mmol) and potassium carbonate (4.8 g, 34.7 mmol). Benzylmercaptan (4.09 mL, 34.7 mmol) was added followed by water (30 mL). Themixture stirred at 25° C. for 4 h. Water (350 mL) was added and theproduct precipitated. The solid was collected by vacuum filtration,rinsed with water (100 mL) and dried in vacuo for 4 h to afford thedesired product, 2-benzylsulfanyl-3-nitro-pyridine (6.6 g, 26.8 mmol,85%), as a yellow powder. ¹H NMR (400 MHz, DMSO-d₆) δ: 4.47 (2H, s),7.21-7.31 (3H, m), 7.40-7.45 (3H, m), 8.58 (1H, dd, J₁=8.6 Hz, J₂=1.7Hz), 8.83 (1H, dd, J₁=4.7 Hz, J₂=1.7 Hz).

b) 3-Nitro-pyridine-2-sulfonyl chloride

2-Benzylsulfanyl-3-nitro-pyridine (6 g, 24.39 mmol) was dissolved indichloromethane (84 mL). Acetic acid (12 mL) and water (24 mL) wereadded. The mixture was chilled to 0° C. With vigorous stirring,1,3-dichloro-5,5-dimethyl-imidazolidine-2,4-dione (14.4 g, 73.17 mmol)was added portionwise as a suspension in dichloromethane (48 mL). Themixture was allowed to slowly warm to 25° C. and continued to stir for16 h. The mixture was poured into 5% aqueous sodium metabisulfitesolution (100 mL) and shaken well. Dichloromethane (200 mL) and 20%aqueous dibasic potassium phosphate solution (100 mL) were added and themixture was shaken well. The layers were separated. The crude desiredproduct, 3-nitro-pyridine-2-sulfonyl chloride, dissolved in the organiclayer was used directly in the next step without further isolation orcharacterization.

c) 3-Nitro-pyridine-2-sulfonic acid amide

The organic layer was chilled to 0° C. and concentrated aqueous ammoniumhydroxide solution (25 mL) was added. The mixture was stirred for 10min. The mixture was washed with 10% aqueous citric acid solution (addeduntil pH<7, ˜200 mL). The organic layer was dried over magnesiumsulfate, filtered and concentrated in vacuo. The resulting solid wastriturated with a 1:1 mixture of hexanes and ethyl acetate (˜15 mL),collected by vacuum filtration and dried in vacuo for 16 h to afford thedesired product, 3-nitro-pyridine-2-sulfonic acid amide (3.1 g, 15.26mmol, 63%), as a beige solid. ¹H NMR (400 MHz, DMSO-d₆) δ: 7.89 (1H, d,J=4.7 Hz), 7.95 (2H, s), 8.48 (1H, dd, J₁=8.6 Hz, J₂=1.6 Hz), 8.92 (1H,d, J=4.7 Hz).

d) 3-Amino-pyridine-2-sulfonic acid amide

3-Nitro-pyridine-2-sulfonic acid amide (3 g, 14.77 mmol) and iron powder(5 g, <10 micron) were suspended in ethanol (100 mL). Saturated aqueousammonium chloride solution (60 mL) was added and the mixture stirred at105° C. for 1.5 h. Upon cooling to 25° C., ethyl acetate (200 mL) wasadded and the mixture was shaken vigorously. The entire mixture wasfiltered through a plug of Celite. The resulting filtrate was dilutedwith ethyl acetate (100 mL) and the layers were separated. The aqueouslayer was back-extracted with ethyl acetate (3×200 mL). The organiclayers were combined, dried over magnesium sulfate, filtered andconcentrated in vacuo to dryness. The resulting solid was trituratedwith a 3:1 mixture of hexanes and ethyl acetate (˜10 mL). The solid wascollected by vacuum filtration and dried in vacuo for 16 h to afford thedesired product, 3-amino-pyridine-2-sulfonic acid amide (1.95 g, 11.27mmol, 76%), as an off-white powder. ¹H NMR (400 MHz, DMSO-d₆) δ: 5.98(2H, bs), 7.19-7.21 (1H, m), 7.32 (2H, bs), 7.79-7.80 (1H, m), 7.90-7.91(1H, m).

e) (1,1-Dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid ethyl ester

3-Amino-pyridine-2-sulfonic acid amide (1.8 g, 10.4 mmol) was dissolvedin 1,4-dioxane (45 mL). Chlorocarbonyl-acetic acid ethyl ester (1.57 mL,12.48 mmol) was added. The mixture was stirred at 90° C. for 1.5 h. Uponcooling, the mixture was diluted with ethyl acetate (300 mL) and washedwith half-saturated aqueous sodium bicarbonate solution (100 mL)followed by saturated aqueous brine solution (100 mL). The organic layerwas dried over magnesium sulfate, filtered and concentrated in vacuo.The resulting oily residue was treated with toluene (36 mL) andtriethylamine (9 mL). The mixture was stirred at 110° C. in a sealedtube for 1.5 h. The residue never completely dissolved. Upon cooling,the mixture was concentrated in vacuo to afford a thick oil. Triturationwith a minimal amount of ethyl acetate (˜5 mL) solidified the product.The solids were collected by vacuum filtration and dried in vacuo for 16h to afford the desired product,(1,1-dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid ethyl ester (0.3 g, 1.11 mmol, 11%), as a white solid. ¹H NMR (400MHz, DMSO-d₆) δ: 1.21 (3H, t, J=7.0 Hz), 3.71 (2H, s), 4.16 (2H,quartet, J=7.1 Hz), 7.70-7.76 (2H, m), 8.62 (1H, dd, J₁=4.0 Hz, J₂=1.5Hz), 12.29 (1H, bs). LC-MS (ESI) calcd for C₁₀H₁₁N₃O₄S 269.05, found270.1 [M+H⁺].

f) (1,1-Dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid sodium salt

(1,1-Dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid ethyl ester (0.25 g, 0.928 mmol) was dissolved in methanol (5 mL)at approximately 60° C. Sodium hydroxide (0.111 g, 2.79 mmol) wasdissolved in water (2.5 mL). Upon cooling to 25° C., the solutions werecombined. The mixture was stirred at 25° C. for 3 h. The mixture wasstored at −40° C. for 16 h. The mixture was concentrated in vacuo toafford the crude product,(1,1-dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid sodium salt (˜0.928 mmol), as a yellow film, which was used in thenext step without further purification.

g)(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one

The crude(1,1-dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-aceticacid sodium salt (˜0.928 mmol),(1S,2R,3S,4R)-3-(4-fluoro-benzylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (0.27 g, 0.928 mmol) andO-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (0.494 g, 1.3 mmol) were combined and dissolved inanhydrous N,N-dimethylformamide (2 mL). The mixture was stirred at 25°C. for 1.5 h. Triethylamine (0.697 mL, 5 mmol) was added and the mixturewas stirred at 50° C. for 16 h. Upon cooling, the mixture was dilutedwith ethyl acetate (100 mL), washed with saturated aqueous ammoniumchloride solution (2×25 mL) and saturated aqueous brine solution (25mL). The organic layer was dried over magnesium sulfate, filtered andconcentrated in vacuo. The resulting solid was triturated with ethylacetate (˜15 mL) and the product solidified. The solid was collected byvacuum filtration and dried in vacuo for 16 h to afford the desiredproduct,(1R,2S,7R,8S)-5-(1,1-dioxo-1,4-dihydro-1λ⁶-thia-2,4,8-triaza-naphthalen-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one(0.03 g, 0.064 mmol, 6.9%), as a white powder. ¹H NMR (400 MHz, DMSO-d₆)δ: 0.92-1.05 (2H, m), 1.18-1.46 (4H, m), 2.26 (1H, d, J=3.1 Hz), 2.36(1H, d, J=9.5 Hz), 2.45 (1H, d, J=3.0 Hz), 3.15 (1H, d, J=9.4 Hz), 4.13(1H, d, J=15.6 Hz), 4.86 (1H, d, J=15.6 Hz), 7.01-7.05 (2H, m),7.16-7.20 (2H, m), 7.39-7.42 (1H, m), 7.50 (1H, dd, J₁=8.6 Hz, J₂=1.6Hz), 8.26 (1H, dd, J₁=4.6 Hz, J₂=1.5 Hz). LC-MS (ESI) calcd forC₂₃H₂₁FN₄O₄S 468.13, found 469.2 [M+H⁺].

Example 66N-{3-[(1R,2S,7R,8S)-3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

a)(1S,2R,3S,4R)-3-(2-Cyclopropyl-ethylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

A 1.4M solution of cyclopropylacetaldehyde in dichloromethane (preparedas described in Example 23a, 3.4 mL, 4.76 mmol) was added to a solutionof (1S,2R,3S,4R)-3-amino-bicyclo[2.2.1]heptane-2-carboxylic acid ethylester (prepared as described in Example 6k, 580 mg, 3.17 mmol) inanhydrous methanol (15 mL) at 25° C. under a nitrogen atmosphere. Afterstirring for 20 min, glacial acetic acid (0.6 mL) was added. Thesolution was cooled to 0° C., sodium triacetoxyborohydride (1.7 g, 7.93mmol) was added, and the resulting mixture was stirred at 25° C. for 20h. The reaction mixture was quenched with saturated aqueous sodiumbicarbonate solution (25 mL) and was extracted with ethyl acetate (3×60mL). The combined organic layers were washed with saturated aqueousbrine solution, dried over sodium sulfate and filtered. The filtrate wasconcentrated in vacuo to afford the desired product,(1S,2R,3S,4R)-3-(2-cyclopropyl-ethylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (676.4 mg, 2.69 mmol, 84.9%), as a yellow oil. LC-MS(ESI) calcd for C₁₅H₂₅NO₂ 251.19, found 252.0 [M+H⁺].

b)(1S,2R,3S,4R)-3-{(2-Cyclopropyl-ethyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester

To a stirred solution of(1S,2R,3S,4R)-3-(2-cyclopropyl-ethylamino)-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester (338.2 mg, 1.35 mmol) in anhydrousN,N-dimethylformamide (10 mL) under a nitrogen atmosphere,(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 493 mg, 1.48 mmol),N-methylmorpholine (0.33 mL, 2.96 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (284 mg,1.48 mmol) were added sequentially. After shaking at 25° C. for 21 h,additional(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-aceticacid (prepared as described in Example 1g, 150 mg, 0.45 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (85 mg, 0.44mmol) were added. After another 26 h, a 1.0M aqueous hydrochloric acidsolution (12 mL) was added and the mixture was extracted with ethylacetate (3×60 mL). The combined organic layers were washed withsaturated aqueous brine solution (15 mL), dried over sodium sulfate,filtered, and concentrated in vacuo to afford the crude product,(1S,2R,3S,4R)-3-{(2-cyclopropyl-ethyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester, as a red oil, which was used in the next step withoutany further purification. LC-MS (ESI) calcd for C₂₅H₃₄N₄O₇S₂ 566.19,found 567.4 [M+H⁺].

c)N-{3-[(1R,2S,7R,8S)-3-(2-Cyclopropyl-ethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

To a solution of the crude(1S,2R,3S,4R)-3-{(2-cyclopropyl-ethyl)-[2-(7-methanesulfonylamino-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-3-yl)-acetyl]-amino}-bicyclo[2.2.1]heptane-2-carboxylicacid ethyl ester in absolute ethanol (11 mL) was added a 21 wt. %solution of sodium ethoxide in ethanol (2.4 mL, 7.41 mmol). Aftershaking at 25° C. for 23 h, additional 21 wt. % solution of sodiumethoxide in ethanol (1.0 mL, 3.09 mmol) was added. After shaking at 25°C. for 6 h, additional 21 wt. % solution of sodium ethoxide in ethanol(1.0 mL, 3.09 mmol) was added and the mixture was shaken for another 24h. The mixture was then acidified with 1.0M aqueous hydrochloric acidsolution (22 mL) and was extracted with ethyl acetate (3×60 mL). Thecombined organic layers were dried over sodium sulfate, filtered, andconcentrated in vacuo. The crude mixture was purified by prep-HPLC[Column Luna 5μ C18 (2) 100 Å AXIA 150×21.2 mm, 5 micron, 30%-95% in 7min @30 mL/min flow rate, 0.05% trifluoroacetic acid inacetonitrile/0.05% trifluoroacetic acid in water] to afford the desiredproduct,N-{3-[(1R,2S,7R,8S)-3-(2-cyclopropyl-ethyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(160.7 mg, 0.309 mmol, 22.9% over two steps), as a solid. ¹H NMR (400MHz, CDCl₃) δ: 0.09-0.15 (2H, m), 0.49-0.54 (2H, m), 0.64-0.73 (1H, m),1.27-1.31 (2H, m), 1.48-1.78 (6H, m), 2.55-2.56 (1H, m), 2.74-2.80 (1H,m), 2.85 (1H, d, J=9.2 Hz), 3.06 (3H, s), 3.10-3.17 (1H, m), 3.57 (1H,d, J=9.2 Hz), 3.80-3.87 (1H, m), 6.99 (1H, s), 7.22-7.25 (1H, m),7.62-7.68 (2H, m). LC-MS (ESI) calcd for C₂₃H₂₈N₄O₆S₂ 520.15, found521.4 [M+H⁺].

Example 67N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6,9-dihydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 168 mg, 0.299 mmol) was dissolvedin dimethyl sulfoxide (15 mL). Reaction buffer was prepared by combininganhydrous D-glucose (3.3 g), GDH-102 (375 mg), and NADP⁺ (570 mg) in a100 mM aqueous potassium phosphate (pH 8.0) solution (700 mL).Lyophilized MCYP-P1C11 (1800 mg, 1500 nmol, 0.84 nmol P450/mg, Codexis,Inc.) was dissolved in a 50 mM aqueous potassium phosphate (pH 8.0)solution (29 mL). The following components were added in order to a 2.8L baffled Fernbach flask: the reaction buffer, followed by theMCYP-P1C11 stock solution, then theN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamidesolution. The vial used to make theN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamidestock solution was washed with 0.1M aqueous potassium phosphate (pH 8.0)solution (6 mL) and this solution was then added to the flask, bringingthe total volume of the reaction to 750 mL. The mixture was thenincubated for 24 h at 30° C. with gentle shaking, followed by freezingat −80° C. Methanol (1.5 L) was added to the thawed reaction mixtureresulting in the formation of a precipitate. The solids were removed asa pellet by centrifugation for 45 min at 10,000 rpm. The supernatant wasconcentrated in vacuo and the residue was purified by prep-HPLC toafford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6,9-dihydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(49.5 mg, 0.086 mmol, 29%), as a white solid. ¹H NMR (500 MHz, DMSO-d₆)δ: 1.21 (1H, dd, J₁=14.7 Hz, J₂=4.8 Hz), 1.46-1.59 (3H, m), 2.44 (1H,s), 2.53 (1H, s), 2.87 (1H, d, J=8.7 Hz), 3.07 (3H, s), 3.39 (1H, d,J=8.5 Hz), 3.80 (1H, d, J=6.0 Hz), 4.42 (1H, d, J=15.5 Hz), 4.96 (1H, d,J=15.7 Hz), 7.16 (2H, t, J=8.7 Hz), 7.32-7.34 (2H, m), 7.52 (1H, dd,J₁=9.0 Hz, J₂=2.3 Hz), 7.59-7.60 (2H, m), 10.23 (1H, s), 14.08 (1H, s),15.03 (1H, bs). LC-MS (ESI) calcd for C₂₅H₂₅FN₄O₇S₂ 576.11, found 577.5[M+H⁺].

Example 68N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl]-methanesulfonamide,L-arginine salt

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 0.280 g, 0.499 mmol) was dissolvedin acetonitrile (5.0 mL). A 0.1M aqueous L-arginine solution (3.0 mL,0.3 mmol) was added, which was followed by addition of a 0.1M solutionof L-arginine in 1-propanol (2.0 mL, 0.2 mmol). After stirring for 6 hat 23° C., the flask was opened to the atmosphere and the suspension wasstirred for 16 h. The solid was collected by filtration and furtherdried in vacuo at 23° C. to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,L-arginine salt, monohydrate (0.257 g, 0.341 mmol, 68%), as acrystalline solid. ¹H NMR (300 MHz, DMSO-d₆) δ: 0.96-1.17 (2H, m), 1.28(1H, app t, J=10.0 Hz), 1.35-1.82 (7H, m), 2.33 (1H, app d, J=3.0 Hz),2.43 (1H, d, J=9.3 Hz), 2.97 (3H, s), 3.00-3.17 (2H, m), 3.23 (1H, d,J=9.3 Hz), 4.21 (1H, d, J=15.3 Hz), 4.94 (1H, d, J=15.3 Hz), 7.04-7.15(3H, m), 7.27 (2H, dd, J=5.7, 8.7 Hz), 7.35 (1H, dd, J=2.5, 8.9 Hz),7.35-7.51 (4H, m), 8.82 (1H, br s), 15.29 (1H, br s). Anal. calcd forC₃₁H₃₉FN₈O₈S₂.H₂O: C, 49.46; H, 5.49; N, 14.88; 0, 19.13; S, 8.52; F,2.52; found: C, 49.49; H, 5.23; N, 14.96; 0, 18.69; S, 8.82; F, 2.81.m.p.=216° C. (DSC).

Example 69N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,L-lysine salt

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 0.090 g, 0.160 mmol) was dissolvedin acetonitrile (2.5 mL). An aqueous solution of L-lysine (0.469 mL of a50 mg/mL solution in water, 0.160 mmol) was added. The solvent wasallowed to evaporate under a flow of nitrogen and ethanol (0.5 mL) wasadded. The mixture was stirred at 35° C. for 2 d, and was then immersedin an ultrasonic bath. Water (0.5 mL) was added, and the mixture wasstirred at 23° C. for 3 d. The solid was collected by filtration andfurther dried in vacuo at 23° C. to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,L-lysine salt, monohydrate (0.070 g, 0.096 mmol, 60%), as a crystallinesolid. ¹H NMR (300 MHz, DMSO-d₆) δ: 0.96-1.15 (2H, m), 1.22-1.76 (10H,m), 2.34 (1H, app d, J=2.7 Hz), 2.43 (1H, d, J=9.3 Hz), 2.74-2.78 (2H,m), 2.97 (3H, s), 3.18-3.29 (1H, m), 4.21 (1H, d, J=15.3 Hz), 4.95 (1H,d, J=15.6 Hz), 7.07-7.18 (3H, m), 7.27 (2H, dd, J=5.7, 8.7 Hz), 7.36(1H, dd, J=2.4, 8.7 Hz), 7.44 (1H, d, J=2.4 Hz), 15.31 (1H, br s). Anal.calcd for C₃₁H₃₉FN₆O₈S₂.H₂O: C, 51.37; H, 5.70; N, 11.59; 0, 19.87; S,8.85; F, 2.62; found: C, 51.13; H, 5.52; N, 11.63; 0, 20.07; S, 9.20; F,2.71. m.p.=200° C. (DSC).

Example 70N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,hemi magnesium salt

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 0.465 g, 0.829 mmol) was dissolvedin acetone (9.0 mL). A 7-8 wt. % solution of magnesium methoxide inmethanol (0.593 mL, 0.414 mmol) was added. The solvent was evaporated,and the residue was then diluted with water (0.9 mL) and acetone (1.8mL). The resulting mixture was stirred at 23° C. for 16 h. The solid wascollected by filtration and further dried in vacuo at 23° C. to affordthe desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide, hemi magnesium salt, trihydrate (0.377 g, 0.602 mmol, 73%),as a crystalline solid. ¹H NMR (300 MHz, DMSO-d₆) δ: 0.96-1.17 (2H, m),1.22-1.58 (4H, m), 2.33 (1H, br s), 2.44 (1H, d, J=9.6 Hz), 2.98 (3H,s), 3.23 (1H, d, J=9.3 Hz), 4.21 (1H, d, J=14.7 Hz), 4.94 (1H, d, J=15.3Hz), 7.03-7.19 (3H, m), 7.21-7.48 (4H, m), 9.81 (1H, br s), 15.35 (1H,br s). Anal. calcd for C₂₅H₂₄N₄O₆FS₂.0.5 Mg.3 H₂O: C, 47.98; H, 4.83; N,8.95; 0, 23.01; S, 10.25; F, 3.04; Mg, 1.94; found: C, 47.66; H, 4.89;N, 8.98; 0, 23.00; S, 11.36; F, 3.09; Mg, 1.82. m.p.=184° C. (DSC).

Example 71N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,sodium salt

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 0.407 g, 0.726 mmol) was suspendedin ethanol (11.0 mL). A 1.0M aqueous sodium hydroxide solution (0.726mL, 0.726 mmol) and water (1.0 mL) were added. The mixture was seededwith a crystal ofN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,sodium salt (produced from a separate batch), and the mixture was thenstirred at 23° C. for 1 d. The solid was collected by filtration andfurther dried in vacuo at 23° C. to afford the desired product,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,sodium salt, hydrate (2.25 molar equiv. water) (0.235 g, 0.377 mmol,52%), as a crystalline solid. ¹H NMR (300 MHz, DMSO-d₆) δ: 0.99-1.11(2H, m), 1.28 (1H, app t, J=10.2 Hz), 1.36-1.53 (3H, m), 2.33 (1H, appd, J=2.7 Hz), 2.42 (1H, d, J=9.3 Hz), 2.97 (3H, s), 3.22 (1H, d, J=9.3Hz), 4.20 (1H, d, J=15.3 Hz), 4.95 (1H, d, J=15.3 Hz), 7.09-7.16 (3H,m), 7.25-7.36 (3H, m), 7.42 (1H, d, J=2.4 Hz), 9.79 (1H, s), 15.32 (1H,s). Anal. calcd for C₂₅H₂₄FN₄NaO₆S₂.2.25 H₂O: C, 48.19; H, 4.61; N,8.99; 0, 21.18; S, 10.29; F, 3.05; Na, 3.69; found: C, 48.14; H, 4.67;N, 8.97; 0, 21.07; S, 10.25; F, 3.13; Na, 3.87. m.p.=182-188° C. (DSC).

Example 72N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,potassium salt

N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide(prepared as described in Example 6, 0.281 g, 0.501 mmol) was dissolvedin methyl ethyl ketone (8.0 mL). A 0.5M aqueous potassium hydroxidesolution (1.0 mL, 0.500 mmol) was added. The solution was seeded withcrystallineN-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,potassium salt (produced from a separate batch), and the resultingmixture was then stirred at 23° C. for 3 h. The solid was collected byfiltration and further dried in vacuo at 23° C. to afford the desiredproduct,N-{3-[(1R,2S,7R,8S)-3-(4-fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-benzo[1,2,4]thiadiazin-7-yl}-methanesulfonamide,potassium salt, hydrate (0.75 molar equiv. water) (0.127 g, 0.207 mmol,41%), as a crystalline solid. ¹H NMR (300 MHz, DMSO-d₆) δ: 0.99-1.11(2H, m), 1.27 (1H, app t, J=10.3 Hz), 1.36-1.54 (3H, m), 2.33 (1H, brs), 2.42 (1H, d, J=9.0 Hz), 2.95 (3H, s), 3.22 (1H, d, J=9.3 Hz), 4.20(1H, d, J=15.3 Hz), 4.96 (1H, d, J=15.6 Hz), 7.09-7.15 (3H, m),7.25-7.34 (3H, m), 7.41 (1H, d, J=2.7 Hz), 9.84 (1H, br s), 15.30 (1H,s). Anal. calcd for C₂₅H₂₄FKN₄O₆S₂.0.75 H₂O: C, 49.05; H, 4.20; N, 9.15;0, 17.64; S, 10.48; F, 3.10; K, 6.39; found: C, 48.82; H, 4.11; N, 9.06;0, 17.35; S, 10.37; F, 3.18; K, 6.75. m.p.=278° C. (DSC).

Biological Testing

The ability of compounds of Formula I to inhibit HCV replication can bedemonstrated in the following in vitro assays.

Compounds were tested for HCV polymerase inhibition. Assays wereperformed in a 96-well streptavidin-coated FlashPlate using 20 nMenzyme, 0.5 μCi of [α-³³P]GTP, 0.6 μM GTP, and 250 nM 5′biotinylatedoligo (rG₁₃)/poly rC in 20 mM Tris-HCl, pH 7.5, 5 mM MgCl₂, 5 mMdithiothreitol, 0.1 g/L bovine serum albumin, and 100 U/mL RNAseinhibitor. The reaction was stopped by aspiration after 75 min at 28° C.and the plate was washed several times. After washing and drying theplate, incorporated radioactivity was counted using a Microbetascintillation counter. IC₅₀ values were calculated relative to theuninhibited control and inhibition data were fitted to a 4-parameterIC₅₀ equation. For very potent inhibitors, the data were fitted to atight binding quadratic equation to obtain IC₅₀ values.

Test results (IC₅₀ values) for compounds of Formula I are summarized inTable 1, wherein ++++ means NS5B polymerase inhibition with IC₅₀ valuesless than 0.02 μM, +++ means IC₅₀ values between 0.02 μM and 0.1 μM, ++means IC₅₀ values between 0.1 μM and 1 μM, and + means IC₅₀ valuesbetween 1 μM and 100 μM. Test results for Example numbers 64 and 65 inTable 1 are EC₅₀ values, wherein **** means HCV replicon inhibition withEC₅₀ values less than 0.02 μM, *** means EC₅₀ values between 0.02 μM and0.1 μM, ** means EC₅₀ values between 0.1 μM and 1 μM, and * means EC₅₀values between 1 μM and 100 μM

TABLE 1 Example # IC₅₀ 1 ++++ 2 ++++ 3 ++ 4 ++++ 5 ++ 6 ++++ 7 ++ 8 ++++9 +++ 10 +++ 11 ++++ 12 ++++ 13 +++ 14 ++++ 15 ++++ 16 ++++ 17 +++ 18+++ 19 + 20 ++ 21 +++ 22 +++ 23 ++++ 24 ++ 25 ++++ 26 ++ 27 + 28 ++ 29++++ 30 ++ 31 +++ 32 +++ 33 + 34 + 35 +++ 36 +++ 37 ++++ 38 +++ 39 + 40+++ 41 +++ 42 +++ 43 +++ 44 ++ 45 +++ 46 ++++ 47 + 48 +++ 49 ++++ 50++++ 51 +++ 52 +++ 53 + 54 ++ 55 ++++ 56 ++++ 57 +++ 58 ++++ 59 ++ 60+++ 61 + 62 +++ 63 ++ 64 ** 65 ** 66 +++ 67 ++++HCV Replicon Assay (Replicon EC₅₀ (μM))

The cell culture component of the assay is performed essentially asdescribed by Bartenschlager et al., Hepatology 2002, 35, 694-703,wherein exponentially growing HCV Huh-7/C24 replicon cells are seeded at4.5×103 cells/well in 96 well plates and 24 hours later are treated withsix point half-log concentration of compound. After 72 hours exposurethe media is discarded from the compound assay plate and the cellmonolayers are lysed by addition of 150 □L lysis mixture (Genospectra)with incubation at 53° C. for 45 minutes. Following incubation, eachlysate is thoroughly mixed and 5 □l (NS3 probe) or 10 □L (GAPDH probe)of each lysate is then transferred to the capture plate and analyzed bybDNA assay.

Branched DNA (bDNA) Assay

Based on provided sequences for NS3 [AJ242652], Genospectra (Fremont,Calif., USA) designed and synthesized probes to these analytes (togetherwith GAPDH). Cellular bDNA analysis is carried out essentially asdescribed in the Genospectra protocol (details in Shyamala, V. et al.,Anal. Biochem. 1999, 266, 140-7), wherein target specific captureextenders, label extenders and blocking probes are added to the captureplate after the addition of 5 or 10 μL cell lysate. After annealingovernight, during which the target RNA is captured to the plate viainteraction with the capture extenders, the plate is washed, and thenamplifier (which binds via the label extenders) and label probe aresequentially added.

After subsequent addition of the chemilumigenic substrate (dioxetan),each plate is read by luminometer (Wallac 1420 Multilabel HTS CounterVictor 2). The luminescence signal is proportional to the amount of mRNApresent in each lysate. In addition to the samples, cell lysate only (noprobe) background controls are also included on each bDNA assay plateand the average signal from these control wells is subtracted from thesample reading prior to analysis. Percent of no drug control isdetermined for both the NS3 and GAPDH signals for each compound also.Percent inhibition is determined for each compound concentration inrelation to the no drug control to calculate the EC₅₀.

Luciferase-Based HCV Replicon Assay Protocol

Exponentially growing HCV Huh-luc/neo-ET replicon cells were seeded at6×10³ cells/well in 96 well assay plate. 24 hours later the cells weretreated with various concentrations of compound in triplicate. After 72hours exposure to the compound the luciferase activity in the wells wasdetermined using Bright-Glo reagent (Promega, Madison, Wis.) with aluminometer (Wallac 1420 Multilabel HTS Counter Victor 2). Thebackground control was replicon cells treated with 100 nM BILN-2061, aninhibitor of the HCV protease. % Inhibition was determined for eachcompound concentration in relation to the negative (no compound) controlto calculate the EC₅₀.

It is to be understood that the foregoing description is exemplary andexplanatory in nature, and is intended to illustrate the invention andits preferred embodiments. Through routine experimentation, the artisanwill recognize apparent modifications and variations that may be madewithout departing from the spirit of the invention.

What is claimed is:
 1. A compound of Formula I

wherein X is N, A is

Ring B is heterocyclyl, optionally substituted by 1-3 R¹ moieties,wherein is R¹ is H, halo, nitro, —CHR⁴—S(O)₂R⁵, —C(S(O)₂R⁵)═CHR⁴—,—NR⁵R⁶, —NR⁴S(O)₂R⁵, or —NR⁴S(O)₂NR⁵R⁶, wherein R⁴, R⁵, and R⁶ areindependently H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C(O)O-(C₁-C₆ alkyl),aryl, or heterocyclyl, or R⁴ and R⁵ or R⁵ and R⁶ combine with theatom(s) to which they are attached to form a 5- or 6-memberedheterocyclyl ring, R² is H, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, -C₁-C₆alkylene(C₃-C₈ cycloalkyl), —C₁-C₆ alkylene(aryl), -C₁-C₆alkylene(heterocyclyl), aryl, or heterocyclyl, Z is —(CR¹³R¹⁴)_(n)—, orO, n is 1 or 2, R⁷ and R⁸ are independently H or C₁-C₈ alkyl, R⁹, R¹⁰,R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are independently H, C₁-C₆ alkyl,hydroxy, or halo, wherein the above alkyl, alkylene, aryl, cycloalkyl,or heterocyclyl moieties provided in R¹, R², R⁴, R⁵, R⁶, R⁷, R⁸, R⁹,R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each optionally andindependently substituted by 1-3 substituents selected from alkylamine,amino, aryl, cycloalkyl, heterocyclyl, C₁-C₆ alkyl, C_(l)-C₆ haloalkyl,C_(l)-C₆ hydroxyalkyl, C_(l)-C₆ alkoxy, C_(l)-C₆ alkylamine, C_(l)-C₆dialkylamine, C₂-C₆ alkenyl, or C₂-C₆ alkynyl, wherein each of which maybe interrupted by one or more hetero atoms, carboxyl, cyano, halo,hydroxy, keto, nitro, —C(O)OH, —C(O)NH₂, —C(O)(C_(l)-C₆ alkylamine),—C(O)(C₁-C₆ dialkylamine), —C(O)₂—(C₁-C₆ alkyl), —C(O)₂—(C₃-C₈cycloalkyl), —C(O)₂-(aryl), —C(O)₂- (heterocyclyl), —C(O)₂—(C₁-C₆alkylene)aryl, —C(O)₂—(C₁-C₆ alkylene)heterocyclyl, —C(O)₂—(C₁-C₆alkylene)cycloalkyl, —C(O)(C₁-C₆ alkyl), —C(O)(C₃-C₈ cycloalkyl),—C(O)(aryl), —C(0)(heterocyclyl), —C(O)(C₁-C₆ alkylene)aryl, —C(O)(C₁-C₆alkylene)heterocyclyl, and —C(O)(C₁-C₆ alkylene)cycloalkyl, wherein eachof the above optional substituents can be further optionally substitutedby 1-5substituents selected from amino, cyano, halo, hydroxy, nitro,C₁-C₆ alkylamine, C₁-C₆ dialkylamine, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆alkenyl, and C₁-C₆ hydroxyalkyl, wherein each alkyl is optionallysubstituted by one or more halo substituents, or a pharmaceuticallyacceptable salt or stereoisomer thereof.
 2. The compound according toclaim 1 wherein R¹ is H, halo, nitro, —CHR⁴—S(O)₂R⁵, —C(S(O)₂R⁵)═CHR⁴—,—NR⁵R⁶, —NR⁴S(O)₂R⁵, or —NR⁴S(O)₂NR⁵R⁶.
 3. The compound according toclaim 2 wherein R¹ is —NR⁴S(O)₂R⁵, and R⁴ and R⁵ are independently H,C₁-C₆ alkyl, or C₃-C₈ cycloalkyl.
 4. The compound according to claim 3wherein R¹ is selected from


5. The compound according to claim 1 wherein R² is selected from


6. The compound according to claim 5 wherein R² is selected from


7. The compound according to claim 6 wherein R² is selected from


8. The compound according to claim 1 wherein R⁷ and R⁸ are H.
 9. Thecompound according to claim 1 wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵,and R¹⁶ are independently selected from


10. The compound according to claim 9 wherein R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are independently selected from


11. The compound according to claim 10 wherein R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are H or hydroxy.
 12. The compound according to claim11 wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are H.
 13. Thecompound according to claim 1 wherein n is
 1. 14. A compound selectedfrom(1R,2S,7R,8S)-5-(7-Bromo-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,N-{3-[(1R,2S,7R,8S)-3-(4-Fluoro-benzyl)-6-hydroxy-4-oxo-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-5-yl]-1,1-dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-7-yl}-methanesulfonamide,(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[2,3-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one,(1R,2S,7R,8S)-5-(1 ,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[4,3-e][1,2,4]thiadiazin-3-yl-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one, and(1R,2S,7R,8S)-5-(1,1-Dioxo-1,4-dihydro-1λ⁶-pyrido[3,2-e][1,2,4]thiadiazin-3-yl)-3-(4-fluoro-benzyl)-6-hydroxy-3-aza-tricyclo[6.2.1.0^(2,7)]undec-5-en-4-one.
 15. Apharmaceutically acceptable composition comprising a compound of claim 1and a pharmaceutically acceptable carrier.
 16. A method for treatinghepatitis C virus infection in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of claim 1, or a pharmaceutically acceptable salt thereof. 17.The method of claim 16 wherein the mammal is a human.
 18. The method ofclaim 16 further comprising administering one or more additionaltherapeutic agents to the mammal.